agradecimentos especiais vão para o Dr. Benjamin Motsh por sua assistência em questões técnicas. Os autores também gostaria de reconhecer o Dr. Marcus Forschner e Birk Müller pelo seu apoio multimédia, Erica Magelky para sua perícia editorial e Lisa Hornung, Dr. Roland Jurgons e Professor Stephan von Hï¿½sten (todos do Franz-Penzoldt-Center, University of Erlangen) pelo seu profissionalismo no manuseio e cuidados com os animais. Agradecemos ao professor Michael Neumaier no Instituto de Química Clínica, Faculdade de Medicina Mannheim, da Universidade de Heidelberg, Alemanha por fornecer células tumorais MC38. O presente trabalho foi realizado em cumprimento dos requisitos para a obtenção do grau &quot;Dr. med.&quot; na Friedrich-Alexander-University Erlangen-Nürnberg (FAU).

O governo de Franconia médio em Baviera, Alemanha, permissão para que os procedimentos descritos concedido. Quaisquer experimentos semelhantes requerem uma autorização prévia das autoridades competentes. Nota: O seguinte manual pode ser usado para grupos previamente discutida de A a C. passos que têm de ser deixado de fora nos grupos A e B são marcadas em conformidade. 1. Preparações <ol><li> Colocar luvas, coloque a almofada de poliestireno sob o microscópio, eo foco da lente na área um pouco acima do teclado. </li><li> Dividir a folha estéril e coloque metade sobre a almofada de poliestireno e a outra metade apenas próximo a ele. </li><li> Esterilizar os instrumentos cirúrgicos e colocá-los em uma folha estéril ao lado do bloco de poliestireno. </li><li> Solte um clipe de papel grande para formar um arco. Vire de cabeça para baixo e pressione-o no bloco de poliestireno na extremidade superior - mesmo ao lado, onde a cabeça do animal vou mentir. </li><li> Colocar o bocal do vaporizador entre os dois membross do arco. (Figura 7) </li><li> Configurar uma lâmpada de aquecimento de cerca de 40 cm do local onde a cabeça do animal vou mentir. Certifique-se o calor ao nível do bloco de poliestireno não exceda 40 ° C. </li><li> Prepara-se uma gaiola separada para os animais operados. </li><li> Prepara-se uma quantidade e volume definido (máximo de 50 ul) de células tumorais num tubo flexível e armazená-lo em gelo. </li></ol> 2. Anestesia <ol><li> Colocar o rato dentro de uma caixa de plexiglas e começar a indução da anestesia de isoflurano alto fluxo (5% de isoflurano a um caudal de 10 L / min). </li><li> Depois de passar pelas diferentes fases da anestesia, remova o mouse na caixa de plexiglas logo depois que entrou na fase da respiração agonia, que pode ser facilmente reconhecido por uma diminuição drástica na taxa respiratória (&lt;20 / min) e suspiros profundos apertando o o corpo inteiro do animal. </li><li> colocar rapidamente o animal de barriga para cima sobre a cortina coberto almofada de poliestireno estéril e continuarde ventilação na isoflurano baixo fluxo, inserindo o focinho do rato no bocal para manter a anestesia. Usar uma fracção inspiratória de 1,8-2,2% a um caudal de cerca de 1 L / min. </li><li> Avaliar a profundidade da anestesia durante a operação por meio do cálculo da frequência respiratória, que é de preferência entre 45 e 60 respirações por minuto. Adaptar a fração isoflurano inspiratória de acordo, se este não é o caso. Nota: As alterações na fração isoflurano inspiratória levar cerca de 60 segundos até que se tornem eficaz. Evite realizar mudanças drásticas rapidamente. Em vez disso, modificar a aplicação do anestésico gradualmente. </li></ol> 3. Operação <ol><li> Desinfectar o tórax e abdômen com um desinfectante adequado e trocar as luvas depois. </li><li> Injectar um volume adaptado de peso de carprofeno (5 mg / kg de peso corporal) na coxa do animal. </li><li> Aperte suavemente a pele abdominal com uma pinça para testar se a profundidade da anestesia é adequada. </li><li&gt; Realizar uma incisão ao longo da linha mediana do xifóide ao mid-abdomen usando tesouras e fórceps. </li><li> Cuidadosamente dissecar a área em torno do xifóide ao expô-la a partir do tecido circundante. </li><li> Colocar uma estadia de sutura através da xifóide (de dentro para fora) e anexar as duas threads ao retentor acima dos animais cabeça, usando a braçadeira. </li><li> Comprima membros do afastador juntos e lentamente introduzir o &quot;U&quot; dicas afastador -shaped ao longo da parede abdominal interna do animal. Nota: Estas medidas expor o fígado para facilitar o acesso aos diferentes lobos. A identificação de lobo lateral mediano e esquerdo do fígado deve agora ser possível. </li><li> Utilize um cotonete de algodão embebido em soro fisiológico e empurre o lobo médio para baixo. Dissecar as ventrais três quartos do ligamento falciforme, que será agora visível entre a superfície do lobo médio e do diafragma. </li><li> Agora, use duas compressas de algodão embebido com solução salina para mudar a medianalóbulo esquerdo e lobo lateral para cima contra o diafragma. </li><li> Visualizar a membrana fina entre o lobo lateral esquerdo e do lobo caudado e dissecá-lo com cuidado. Nota: Ao executar este protocolo em animais do grupo A, salte para a etapa 3.18 neste momento. </li><li> Coloque uma ligadura tamanho 4-0 na diagonal ao longo da base do lobo lateral esquerdo. </li><li> Em seguida, use o cotonete para devolver o lobo lateral esquerdo para sua posição original. </li><li> Cuidadosamente amarrar a ligadura o mais próximo possível da base do lobo possível e avaliar a mudança de cor no lobo para testar o fornecimento de sangue adequadamente interrompido. </li><li> Ressecção do lobo lateral esquerdo, seguindo a linha de base do lóbulo e observe o peso do lobo ressecado. Nota: Ao executar este protocolo em animais do grupo B, pule para o passo 3.18 neste momento. </li><li> Coloque uma segunda ligadura entre coto do lobo lateral esquerdo e base do lobo mediano. </li><li> Reposicionar o lobo mediano como well e amarrar a ligadura. Mais uma vez, para avaliar a alteração de cor no lobo a testar para fornecimento de sangue adequadamente interrompida. </li><li> Ressecção do lobo mediano e anote o seu peso. </li><li> Ligar a seringa de 1 ml para a agulha 30 G e preenchê-lo com as células do tumor a partir do tubo flexível sem inclinar a seringa, em qualquer momento. </li><li> Use os cotonetes para mover as alças intestinais à esquerda do animal a fim de expor o lóbulo direito inferior. </li><li> Inserir a seringa carregada de células para dentro do dispositivo &quot;de terceira mão&quot; em um ângulo de 30 ° com a vertical. </li><li> Mova cuidadosamente o dispositivo ao lado do rato com a agulha logo acima do lóbulo direito inferior. </li><li> Lentamente avanço a seringa dentro do dispositivo de terceira mão, até que a ponta da agulha se encontra na parte central do lóbulo direito inferior. </li><li> Injectar o volume inteiro na parte central do lobo ao longo de um período de 30-45 segundos. </li><li> Comprimir o local da injecção, durante pelo menos três minutos até a hemorragia parou. </li&gt; <li> Retirar a sutura estadia eo afastador. </li><li> Feche a fáscia com um 5-0 sutura reabsorvível usando nós botão. </li><li> Fechar a pele com uma sutura 4-0, não reabsorvível usando nós botão. </li><li> Comece a ventilação do mouse no alto fluxo de oxigênio durante cerca de 1 min. </li></ol> 4. Pós-op procedimento <ol><li> Colocar o animal num ambiente aquecido (aproximadamente 40 ° C) durante a meia hora seguinte para assegurar a recuperação adequada. </li><li> Admix água de beber do animal com 5 mg / ml de metamizol para pós-operatório de 72 h. </li><li> Examinar a integridade das suturas de perto durante pelo menos três dias após o procedimento. </li></ol> 5. Período de Observação e Eutanásia <ol><li> Medir o peso do animal por dia, durante um total de 14 dias. Pontuação-los quanto ao seu bem-estar e limitações respectivamente. </li><li> Após 14 dias, anestesiar o seguinte animais passos 2.1 e 2.2 do protocolo operatório umnd prosseguir com o protocolo da instituição para a eutanásia dos animais. </li><li> Realizar uma laparotomia mediana como explicado no passo 3.4. Para facilitar o acesso ao abdômen, estender a incisão 1-1,5 cm caudalmente. Insira o afastador como apontado em 3,7. </li><li> Dissecar ao longo do resto do ligamento falciforme e cortar a veia cava inferior do mesmo modo que sai do fígado cranialmente. </li><li> Separa-se o fígado a partir do diafragma, o diafragma, agarrando com os fórceps e dissecando bruscamente para dentro do espaço entre o fígado e tecido muscular. </li><li> Levante o tecido do fígado mobilizados fora do retroperitônio e dissecá-lo fora das estruturas remanescentes que ainda está ligado: tecido adiposo retroperitoneal, a veia cava inferior ea veia portal. </li><li> Inspeccionar o fígado após a extracção para qualquer tecido adicional, que seria falsamente contribuir para o seu tamanho e o peso real. Um fígado e extraiu-se o seu tumor são apresentadas na Figura 6. </li><li> Dissecar o tumor fora da inferior lóbulo direito. </li><li> Medir o tamanho e o peso de ambos o tumor e o parênquima do fígado. </li><li> Conservar amostras adicionais de tecidos do peritônio, linfonodos ou outros órgãos, conforme necessário para análise de tecido </li></ol>

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The authors have nothing to disclose.

Experimentos anteriores realizando a cirurgia em roedores foram capazes de identificar certas variáveis ​​que poderiam servir como fontes de viés. A fim de obter resultados confiáveis ​​e válidos, considere as seguintes precauções. Rotina jejum pré-operatório pode levar a esteatose hepática 12, que pode inibir a regeneração do fígado 13,14. Portanto, não é recomendado. A maior actividade mitótica de hepatócitos varia ao longo do dia 15.</sup...

O câncer colorretal (CRC) é responsável por cerca de 9% de todos os tumores malignos. É o terceiro cancro mais comum, tanto os EUA e em todo o mundo. As taxas de mortalidade globais de gama CRC de 300.000 para mais de 500.000 por ano 1. Vinte por cento dos pacientes sofrem de metástases hepáticas após a descoberta do seu tumor colo-rectal. Metástases ressecáveis ​​são normalmente tratados por um 2,3 parcial ressecção hepática. Técnicas cirúrgicas melhoradas, novas estratégias multimodais e novas definições de metástases ressecáveis ​​tornar a terapia de uma ressecção parcial do fígado possível para um número crescente de pacientes 4. Recorrência de tumores hepáticos secundários, no entanto, é uma seqüelas clínica desafiadora na cirurgia gastrointestinal moderna. Os pacientes com CRC submetidos à ressecção de metástases hepáticas têm uma chance de 30 a 50% de desenvolver um novo tumor em seu fígado remanescente 5. Portanto, há uma necessidade de mais pesquisas sobre omecanismos envolvidos na recorrência de metástases hepáticas. A ressecção hepática de cerca de 70% é normalmente compensada dentro de algumas semanas por o tecido hepático remanescente. Esta regeneração envolve vários mecanismos, incluindo citocinas, como interleucina 6 (IL-6), factor de necrose tumoral alfa (TNF-α), factor de crescimento de hepatócitos (HGF), factor de crescimento transformante beta (TGF-β), factor de crescimento endotelial vascular (VEGF ), metaloproteases de matriz (MMP-2 e MMP-9) e CXC-quimiocinas 6-11. Estas substâncias apoiar a regeneração hepática e também pode ser responsável pelas altas taxas de recorrência de tumores malignos do fígado primário e secundário, induzindo o crescimento dos depósitos de células tumorais pequenas no fígado restante que não são detectados pela imagem latente clínica de rotina. Este causalidade não foi provado até agora. A hipótese que se segue foi estabelecida. Após a ressecção parcial do fígado, os fatores de proliferação que são responsáveis ​​por ao vivoR hipertrofia pode também induzir o crescimento de células de tumor previamente desconhecidas no fígado. Um modelo do rato foi projetado que combinou as técnicas de ressecção hepática e indução do tumor. Trinta ratinhos nu / nu-foxn1nu atímicos foram divididos em três grupos de dez animais cada. Cada uma delas foi tratada com qualquer uma laparotomia sozinho (Grupo A), a ressecção hepática 30% (Grupo B) ou a ressecção hepática de 70% (Grupo C). Os animais em todos os grupos subsequentemente receberam uma injecção de células de tumor numa parte restante definido do fígado, para simular células tumorais dormentes. Animais foram observados durante duas semanas e, em seguida, avaliada para o crescimento tumoral e hipertrofia do fígado. O objectivo era criar um modelo que poderia ser utilizado para pesquisar factores moleculares e patogénicos que podem desempenhar um papel na formação de tumores após a hepatectomia. Este método pode ser útil na avaliação: a origem dos factores endócrinos envolvidas na regeneração do fígado; Os mecanismos responsáveis ​​por tum intra-hepáticaou o crescimento após a ressecção hepática; e o volume ressecção hepática necessária para a indução do crescimento de tumores intra-hepática. O método a seguir só foi realizada em animais, porque eles prometem contribuir para a compreensão dos princípios biológicos fundamentais e para o desenvolvimento do conhecimento que pode ser esperado para beneficiar os seres humanos pela melhoria opções de tratamento. Devido aos mecanismos envolvidos nestes assuntos, que teve de ser examinada in vivo, como métodos in vitro podem não fornecer uma representação realista da patologia humana. Estas investigações podem levar à descoberta de metas relevantes para as opções de tratamento profilático para diminuir a recorrência do tumor.

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			<td height="20" style="height:20px;width:319px;">Equipment</td>
			<td style="width:217px;"></td>
			<td style="width:143px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Operation Microscope</td>
			<td style="width:217px;">Zeiss</td>
			<td style="width:143px;">OPMI-1 FC S21</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Induction Cage (Plexiglas Box)</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000132</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Flowmeter + Connection Kit</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000008</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">UNO Vaporizer Sigma Delta</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000002</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Key Filler for Anesthetic</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000010</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Activated Charcoal Filter Adsorber</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000140</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Gas Exhaust Unit</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000118</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Face Mask for mouse</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000065</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Vaporizer Stand</td>
			<td style="width:217px;">UNO BV, Netherlands</td>
			<td style="width:143px;">180000006</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Heat lamp</td>
			<td style="width:217px;">Physitemp Instruments</td>
			<td style="width:143px;">HL-1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Styrofoam Pad</td>
			<td style="width:217px;">RAYHER</td>
			<td style="width:143px;">30074000&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Third Hand Tool</td>
			<td style="width:217px;">TOOLCRAFT&#160;</td>
			<td style="width:143px;">ZD-10F</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Precision Scales</td>
			<td style="width:217px;">Kern</td>
			<td style="width:143px;">EW 220-3NM</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Scales&#160;</td>
			<td style="width:217px;">Kern</td>
			<td style="width:143px;">EMB 500-1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Sliding Caliper</td>
			<td style="width:217px;">MIB</td>
			<td style="width:143px;">MIB 82026100</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Microdissection forceps</td>
			<td style="width:217px;">Braun/Aesculap</td>
			<td style="width:143px;">BD195R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Microdissection scissors</td>
			<td style="width:217px;">Braun/Aesculap</td>
			<td style="width:143px;">FD100R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Microdissection needle holder</td>
			<td style="width:217px;">Braun/Aesculap</td>
			<td>BM563R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Retractor</td>
			<td style="width:217px;">Fine Science Tools (F.S.T.)</td>
			<td style="width:143px;">No. 17001-0</td>
			<td>Type: Bowmann</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Clamp</td>
			<td style="width:217px;">Braun/Aesculap</td>
			<td style="width:143px;">BJ002R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Expendable Items</td>
			<td style="width:217px;"></td>
			<td style="width:143px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td colspan="4" height="20" style="height:20px;width:845px;">(NOTE: Quantities are per animal and procedure)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Foliodrape sterile cover (45x75cm)</td>
			<td style="width:217px;">Hartmann</td>
			<td style="width:143px;">2775001</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Sterile Cotton Swabs (2x)</td>
			<td style="width:217px;">Hartmann</td>
			<td style="width:143px;">4700151</td>
			<td>Peha</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Sterile fluid (0,9%NaCl)</td>
			<td style="width:217px;">Braun</td>
			<td style="width:143px;">3570310</td>
			<td>PZN=04454809</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Disinfectant (Softasept - 250ml)</td>
			<td style="width:217px;">Braun</td>
			<td style="width:143px;">3887138</td>
			<td>PZN=0762008808505018</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">2 x 1ml syringe (Injekt-F )</td>
			<td style="width:217px;">Braun</td>
			<td>9166017V</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">26G canula (Sterican) - for Carprofen injection</td>
			<td style="width:217px;">Braun</td>
			<td>4665457</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">30G canula (Sterican)&#160; - for Tumor injection</td>
			<td style="width:217px;">Braun</td>
			<td>4656300</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Caprofen (=Rimadyl)</td>
			<td style="width:217px;">Pfizer</td>
			<td style="width:143px;">QM01AE91</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Metamizole (=Novaminsulfon)</td>
			<td style="width:217px;">Ratiopharm</td>
			<td style="width:143px;">16543.00.00</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">4-0 Vicryl suture</td>
			<td style="width:217px;">Ethicon</td>
			<td style="width:143px;">J835G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">5-0 Prolene suture</td>
			<td style="width:217px;">Ethicon</td>
			<td style="width:143px;">8618G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SafeLock Flex-Tube 1.5mL</td>
			<td style="width:217px;">Eppendorf&#160;</td>
			<td style="width:143px;">22363778</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">4x4 Gauze Sponge</td>
			<td style="width:217px;">Kendall/Covidien&#160;</td>
			<td style="width:143px;">UPC: 728795135355&#160;</td>
			<td>ASIN: B005BFQTWM&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Large paperclip</td>
			<td style="width:217px;">ACCO</td>
			<td style="width:143px;">A7072510G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Animals</td>
			<td style="width:217px;"></td>
			<td style="width:143px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:319px;">Female athymic nude-foxn1nu/nu</td>
			<td style="width:217px;">Harlan Laboratories B.V.</td>
			<td style="width:143px;">Code 069</td>
			<td></td>
		</tr>
	</tbody>
</table>

the influence of liver resection on intrahepatic tumor growth

A alta incidência de recorrência do tumor após a ressecção de lesões hepáticas metastáticas permanece um problema não resolvido. depósitos de células de tumor pequeno, que não são detectáveis ​​por imagem clínica de rotina, pode ser estimulada por factores de regeneração hepáticas após a ressecção hepática. Não está inteiramente claro, no entanto, que fatores são cruciais para a recorrência do tumor. O modelo de ratinho apresentada pode ser útil para explorar os mecanismos que desempenham um papel no desenvolvimento de lesões malignas recorrentes após a ressecção hepática. O modelo combina as técnicas fáceis de realizar e reprodutível de quantidades definidas de remoção de tecido do fígado e a indução de tumor (por injecção) em ratinhos. Os animais foram tratados com uma única laparotomia, uma ressecção hepática de 30%, ou de uma ressecção hepática de 70%. Todos os animais receberam, subsequentemente, uma injecção das células tumorais no tecido do fígado remanescente. Depois de duas semanas de observação, os fígados e os tumores foram avaliados quanto a tamanho e peso eexaminada por imuno-histoquímica. Após a ressecção hepática de 70%, o volume do tumor e peso foram significativamente aumentados em comparação com uma laparotomia sozinhos (p &lt;0,05). Adicionalmente, imuno-histoquímica (Ki67) mostrou um aumento na taxa de proliferação do tumor no grupo de ressecção (p &lt;0,05). Estes resultados demonstram a influência dos mecanismos de regeneração hepática no crescimento do tumor intra-hepática. Combinado com métodos como workup histológica ou análise de RNA, o modelo de rato descrito poderia servir como base para um exame atento dos diferentes fatores envolvidos no crescimento tumoral e recorrência da doença metastática no fígado. Um número considerável de variáveis, como o tempo de observação pós-operatória, a linha celular utilizada para injecção ou o tempo de injeção e ressecção hepática oferecer vários ângulos ao explorar uma questão específica no contexto de metástases pós-hepatectomia. As limitações deste procedimento são a aurização para realizar o procedimento em animais, o acesso a uma instalação de testes em animais adequados e aquisição de certos equipamentos.

Em nosso experimento específico, que incluiu 30 ratos nude-foxn1nu / nu atímicos. Eles receberam uma laparotomia mediana, as injecções de células de tumor de 500.000 células tumorais MC38 (dissolvida em 50 ul de solução salina), e foram tratados subsequentemente quer com uma ressecção hepática de 70%, uma ressecção hepática de 30%, ou mais nenhuma intervenção. Após 14 dias, a regeneração quase completa do f?...

Watch this Scientific Journal Video about The Influence of Liver Resection on Intrahepatic Tumor Growth at JoVE.com

The Influence of Liver Resection on Intrahepatic Tumor Growth

A high incidence of tumor recurrence after resection of liver metastases remains an unsolved problem. The illustrated mouse model may be useful to investigate the reasons for such recurrences. It combines a liver resection model with intrahepatic tumor cell injection for the first time.

A Influência do fígado ressecção no crescimento do tumor intra-hepática

The high incidence of tumor recurrence after resection of metastatic liver lesions remains an unsolved problem. Small tumor cell deposits, which are not ...

the-influence-of-liver-resection-on-intrahepatic-tumor-growth

Research

JoVE Journal

Medicine

9.4K visualizações.  Friedrich-Alexander-University Erlangen-Nürnberg (FAU). A high incidence of tumor recurrence after resection of liver metastases remains an unsolved problem. The illustrated mouse model may be useful to investigate the reasons for such recurrences. It combines a liver resection model with intrahepatic tumor cell injection for the first time. 

Vídeo: The Influence of Liver Resection on Intrahepatic Tumor Growth

An In Vitro System to Study Tumor Dormancy and the Switch to Metastatic Growth

Recurrence of breast cancer often follows a long latent period in which there are no signs of cancer, and metastases may not become clinically apparent until many years after removal of the primary tumor and adjuvant therapy. A likely explanation of this phenomenon is that tumor cells have seeded metastatic sites, are resistant to conventional therapies, and remain dormant for long periods of time 1-4.
The existence of dormant cancer cells at secondary sites has been described previously as quiescent solitary cells that neither proliferate nor undergo apoptosis 5-7. Moreover, these solitary cells has been shown to disseminate from the primary tumor at an early stage of disease progression 8-10 and reside growth-arrested in the patients' bone marrow, blood and lymph nodes 1,4,11. Therefore, understanding mechanisms that regulate dormancy or the switch to a proliferative state is critical for discovering novel targets and interventions to prevent disease recurrence. However, unraveling the mechanisms regulating the switch from tumor dormancy to metastatic growth has been hampered by the lack of available model systems.
in vivo and ex vivo model systems to study metastatic progression of tumor cells have been described previously 1,12-14. However these model systems have not provided in real time and in a high throughput manner mechanistic insights into what triggers the emergence of solitary dormant tumor cells to proliferate as metastatic disease. We have recently developed a 3D in vitro system to model the in vivo growth characteristics of cells that exhibit either dormant (D2.OR, MCF7, K7M2-AS.46) or proliferative (D2A1, MDA-MB-231, K7M2) metastatic behavior in vivo . We demonstrated that tumor cells that exhibit dormancy in vivo at a metastatic site remain quiescent when cultured in a 3-dimension (3D) basement membrane extract (BME), whereas cells highly metastatic in vivo readily proliferate in 3D culture after variable, but relatively short periods of quiescence. Importantly by utilizing the 3D in vitro model system we demonstrated for the first time that the ECM composition plays an important role in regulating whether dormant tumor cells will switch to a proliferative state and have confirmed this in in vivo studies15-17. Hence, the model system described in this report provides an in vitro method to model tumor dormancy and study the transition to proliferative growth induced by the microenvironment.

An In Vitro System to Study Tumor Dormancy and the Switch to Metastatic Growth

A modified 3-D in vitro system is presented in which growth characteristics of several tumor cell lines in reconstituted basement membrane correlate with the dormant or proliferative behavior of the tumor cells at a metastatic secondary site in vivo.

Um In Vitro Sistema para Estudo de dormência Tumor ea Mude para o crescimento metastático

Recurrence of breast cancer often follows a long latent period in which there are no signs of cancer, and metastases may not become clinically apparent ...

Medicina

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

A Novel de alta resolução<em&gt; In vivo</em&gt; Técnica de Imagem para estudar a resposta dinâmica de estruturas intracranianas ao crescimento do tumor e Terapêutica

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

Orthotopic Injection of Breast Cancer Cells into the Mammary Fat Pad of Mice to Study Tumor Growth.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the functions of proteins involved in oncogenic progression or help to discover new tumor suppressors. In addition, injecting cancer cells into mice with different genotypes might provide a better understanding of the importance of the stromal compartment. Many models may be useful to investigate certain aspects of disease progression but do not recapitulate the entire cancerous process. In contrast, breast cancer cells engraftment to the mammary fat pad of mice better recapitulates the location of the disease and presence of the proper stromal compartment and therefore better mimics human cancerous disease. In this article, we describe how to implant breast cancer cells into mice orthotopically and explain how to collect tissues to analyse the tumor milieu and metastasis to distant organs. Using this model, many aspects (growth, angiogenesis, and metastasis) of cancer can be investigated simply by providing a proper environment for tumor cells to grow.

Cancer is a complex disease that is influenced by the tissue surrounding the tumor as well as local pro- and anti-inflammatory mediators. Therefore, orthotropic injection models, rather than subcutaneous models may be useful to study cancer progression in a manner that better mimics human pathology.

Injeção Orthotopic de células cancerosas da mama na mamária Fat Pad de Ratos para o Estudo de crescimento tumoral.

Breast cancer growth can be studied in mice using a plethora of models. Genetic manipulation of breast cancer cells may provide insights into the ...

Establishment of a Human Multiple Myeloma Xenograft Model in the Chicken to Study Tumor Growth, Invasion and Angiogenesis

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the lack of the bone microenvironment and auto/paracrine growth factors human MM cells are difficult to cultivate. Therefore, there is an urgent need to establish proper in vitro and in vivo culture systems to study the action of novel therapeutics on human MM cells. Here we present a model to grow human multiple myeloma cells in a complex 3D environment in vitro and in vivo. MM cell lines OPM-2 and RPMI-8226 were transfected to express the transgene GFP and were cultivated in the presence of human mesenchymal cells and collagen type-I matrix as three-dimensional spheroids. In addition, spheroids were grafted on the chorioallantoic membrane (CAM) of chicken embryos and tumor growth was monitored by stereo fluorescence microscopy. Both models allow the study of novel therapeutic drugs in a complex 3D environment and the quantification of the tumor cell mass after homogenization of grafts in a transgene-specific GFP-ELISA. Moreover, angiogenic responses of the host and invasion of tumor cells into the subjacent host tissue can be monitored daily by a stereo microscope and analyzed by immunohistochemical staining against human tumor cells (Ki-67, CD138, Vimentin) or host mural cells covering blood vessels (desmin/ASMA).
In conclusion, the onplant system allows studying MM cell growth and angiogenesis in a complex 3D environment and enables screening for novel therapeutic compounds targeting survival and proliferation of MM cells.

Human multiple myeloma (MM) cells require the supportive microenvironment of mesenchymal cells and extracellular matrix components for survival and proliferation. We established an in vivo chicken embryo model with engrafted human myeloma and mesenchymal cells to study effects of cancer drugs on tumor growth, invasion and angiogenesis.

Criação de um Modelo de Xenoenxerto Humano mieloma múltiplo na galinha para estudar o crescimento tumoral, invasão e angiogênese

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the ...

Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

Toxicological evaluation is crucial for understanding the effects of chemicals on living organisms in basic and applied biological science fields. A non-mammalian soil round worm, Caenorhabditis elegans, is a valuable model organism for toxicology studies due to its convenience and lack of animal ethics issues compared with mammalian animal systems. In this protocol, a detailed procedure of toxicological evaluation of chemicals in C. elegans is described. A clinical anticancer drug, etoposide, which targets human topoisomerase II and inhibits DNA replication of human cancer cells, was selected as a model testing chemical. Age-synchronized C. elegans eggs were exposed to either dimethyl sulfoxide (DMSO) or etoposide, and then the growth of C. elegans was monitored every day for 4 days by the stereo microscope observation. The total number of eggs laid from C. elegans treated with DMSO or etoposide was also counted by using the stereo microscope. Etoposide treatment significantly affected the growth and reproduction of C. elegans. By comparison of the total number of eggs laid from worms with different treatment periods of chemicals, it can be decided that the reproductive toxicity of chemicals on C. elegans reproduction is reversible or irreversible. These protocols may be helpful for both the development of various drugs and risk assessment of environmental toxicants.

Measuring the Effect of Chemicals on the Growth and Reproduction of Caenorhabditis elegans

A basic protocol to evaluate the toxicity of chemicals in a model animal, Caenorhabditis elegans, is described. The method is convenient and useful for the development of pharmaceuticals as well as for the risk assessment of various environmental pollutants.

Medir o efeito de produtos químicos sobre o crescimento e a reprodução de Caenorhabditis elegans

Toxicological evaluation is crucial for understanding the effects of chemicals on living organisms in basic and applied biological science fields. A ...

Transaxillary First Rib Resection for Treatment of the Thoracic Outlet Syndrome

Thoracic outlet syndrome (TOS) is a common disorder that causes a significant loss of productivity. The transaxillary first rib resection (TFRR) protocol has been used for the decompression of trapped neurovascular structures in the TOS. Among the other surgical procedures, the advantage of the TFRR is that it has the smallest rate of recurrence and better cosmetic outcomes. The disadvantage of TFRR is that it provides a narrow, and deep working corridor that makes obtaining vascular control challenging.

Here, we present a protocol of the transaxillary resection of the first rib for treatment of thoracic outlet syndrome caused by compression of the brachial plexus, subclavian vein and artery.

Ressecção da Primeira Costela Transaxilar para Tratamento da Síndrome do Desfiladeiro Torácico

Thoracic outlet syndrome (TOS) is a common disorder that causes a significant loss of productivity. The transaxillary first rib resection (TFRR) protocol ...

Modified Posterior Vertebral Column Resection for Patients with Thoracolumbar Kyphotic Deformity

Old compression vertebrae fracture or congenital kyphoscoliosis with abnormal vertebral body development and other diseases that invade the spine may cause severe thoracolumbar kyphotic deformity, often accompanied by intractable low back pain or compression of the spinal cord, leading to severe neurological symptoms or even paralysis. If conservative treatment cannot relieve the symptoms or correct the deformities, surgical treatment is usually needed. For severe kyphotic deformity, reconstruction of the physiological curvature and rigid fixation determine the prognosis of the patients. Osteotomy and orthopedics are the standard procedure for deformities with severe compression of the front and middle column, but the trauma to the patients is high, with a long operation time and massive blood loss. To avoid these disadvantages, we have developed a modified technique to remove the diseased vertebra unilaterally. In this technique, we use a modified trephine to resect the vertebral columns like in the pedicle screw technique by adding a locking instrument that can restrict the trephine to lower the risk of osteotomy and shorten the surgery time and blood loss.

We describe a modified technique for resecting the posterior vertebral column unilaterally based on a modified trephine for patients with thoracolumbar kyphotic deformity.

Ressecção da Coluna Vertebral Posterior Modificada para Pacientes com Deformidade Cifótica Toracolombar

Old compression vertebrae fracture or congenital kyphoscoliosis with abnormal vertebral body development and other diseases that invade the spine may ...

An Apical Resection Model in the Adult Xenopus tropicalis Heart

It is known that in adult mammals, the heart has lost its regenerative capacity, making heart failure one of the leading causes of death worldwide. Previous research has demonstrated the regenerative ability of the heart of the adult Xenopus tropicalis, an anuran amphibian with a diploid genome and a close evolutionary relationship with mammals. Additionally, studies have shown that following ventricular apex resection, the heart can regenerate without scarring in X. tropicalis. Consequently, these previous results suggest that X. tropicalis is an appropriate alternative vertebrate model for the study of adult heart regeneration. A surgical model of cardiac regeneration in the adult X. tropicalis is presented herein. Briefly, the frogs were anesthetized and fixed; then, a small incision was made with iridectomy scissors, penetrating the skin and pericardium. Gentle pressure was applied to the ventricle, and the apex of the ventricle was then cut out with scissors. Cardiac injury and regeneration were confirmed by histology at 7-30 days post resection (dpr). This protocol established an apical resection model in adult X. tropicalis, which&#160;can be employed to elucidate the mechanisms of adult heart regeneration.

An Apical Resection Model in the Adult Xenopus tropicalis Heart

Xenopus tropicalis is an ideal model for regenerative research as many of its organs possess a remarkable regenerative capacity. Here, we present a method for constructing a heart injury model in X. tropicalis via apex resection.

Modelo de Ressecção Apical no Coração de Xenopus tropicalis Adulto

It is known that in adult mammals, the heart has lost its regenerative capacity, making heart failure one of the leading causes of death worldwide. ...

Microscopic Cyst Resection for the Treatment of Patients Diagnosed with Epididymal Cyst

Epididymal cysts mostly occur in men aged 20-40 years old. Previous reports have covered concerns about postoperative complexes, including postoperative asoedema, hematoma, sustaining pain, and seminal tract obstruction in patients who have undertaken nonmicroscopic epididymal cyst resection or epididymal resection. Nonmicroscopic epididymal cyst surgery is suggested for patients with childbirth plans as a precaution. The treatment of male epididymal cysts via microtechnology is obviously a beneficial option; we took the lead in carrying out microscopic epididymal exploration and cyst resection surgery in China. From September 2017 to April 2021, 41 young and middle-aged male patients diagnosed with epididymal cysts underwent microtechnology treatment in a program titled &#34;microscopic epididymal exploration and cystectomy&#34;. The postoperative follow-up lasted for 3-50 months. The results confirmed that, as microscopic manipulation largely improved visualization of the subtle tissue structures of the epididymis, the cyst could be clearly dissected apart and completely removed intact under the microscope. Bleeding during the operation was significantly reduced (2-3 mL) and wound drainage was not required. According to follow-up data, microscopic treatment significantly reduced the incidence of postoperative scrotal hematoma, edema, and long-term postoperative pain, thereby promising a higher surgical success rate as well as recurrence prevention. Besides, preliminary experience and reflection suggest that microscopic epididymal exploration and cystectomy provide efficient preservation of the epididymal patency through refined treatment, while a better prognosis can be achieved. We recommend that surgery be carried out before the epididymal cyst develops to 0.8 cm in diameter, for fear that a larger epididymal cyst (&#62;0.9 cm in diameter) could cause the complete destruction of all tubules of the ipsilateral epididymis - a more severe case with damage to the testicular output network.

Here, we present a protocol to treat patients diagnosed with an epididymal cyst using microscopic cyst resection. Based on microscopic manipulation, a largely improved visualization of the subtle tissue structures of the epididymis was obtained, so the cyst could be clearly dissected apart and completely removed intact.

Ressecção de Cisto Microscópico para Tratamento de Pacientes Diagnosticados com Cisto Epididimal

Epididymal cysts mostly occur in men aged 20-40 years old. Previous reports have covered concerns about postoperative complexes, including postoperative ...

Análise quantitativa da distribuição da rigidez hepática

A Three-Dimensional Digital Model for Early Diagnosis of Hepatic Fibrosis Based on Magnetic Resonance Elastography

Hepatic fibrosis is an early stage of liver cirrhosis, and there are no better non-invasive and convenient methods for the detection and evaluation of the disease. Despite the good progress made with the liver stiffness map (LSM) based on magnetic resonance elastography (MRE), there are still some limitations that need to be overcome, including manual focus determination, manual selection of regions of interest (ROIs), and discontinuous LSM data without structural information, which makes it impossible to evaluate the liver as a whole. In this study, we propose a novel three-dimensional (3D) digital model for the early diagnosis of hepatic fibrosis based on MRE.
MRE is a non-invasive imaging technique that employs magnetic resonance imaging (MRI) to measure the liver stiffness at the scanning site through human-computer interaction. Studies have indicated a significant positive correlation between the LSM obtained through MRE and the degree of hepatic fibrosis. However, for clinical purposes, a comprehensive and precise quantification of the degree of hepatic fibrosis is necessary. To address this, the concept of Liver Stiffness Distribution (LSD) was proposed in this study, which refers to the 3D stiffness volume of each liver voxel obtained by the alignment of 3D liver tissue images and MRE indicators. This provides a more effective clinical tool for the diagnosis and treatment of hepatic fibrosis.

Autor em destaque: Avançando no diagnóstico de fibrose hepática usando elastografia por ressonância magnética e IA 

The objective of this study was to develop a novel three-dimensional digital model for the early diagnosis of hepatic fibrosis, which includes the stiffness of each voxel in the patient's liver and can thus, be used to calculate the distribution ratio of the patient's liver at different fibrosis stages.

Modelo digital tridimensional para diagnóstico precoce de fibrose hepática baseado em elastografia por ressonância magnética

Hepatic fibrosis is an early stage of liver cirrhosis, and there are no better non-invasive and convenient methods for the detection and evaluation of the ...