Name: intranasal delivery of therapeutic stem cells to glioblastoma in a mouse model
Uploaded: 2017-06-04T15:00:00
Description: Watch this Scientific Journal Video about Intranasal Delivery of Therapeutic Stem Cells to Glioblastoma in a Mouse Model at JoVE.com

Dette arbejde blev støttet af NIH R01NS087990 (MSL, IVB).

Alle dyreprocedurer skal godkendes af Institutional Animal Care and Use Committee (IACUC) eller tilsvarende. Hvis der er nogen usikkerhed om de specifikke procedurer, der er beskrevet heri, skal du ikke fortsætte. Afklare med institutionens IACUC og et udpeget veterinærmedlem. 1. Sikring af dyrkning af dyrkede celler og følge principper for aseptiske teknikker <ol><li> Følg standard god laboratoriepraksis i alle cellekulturprocedurer og IACUC-krav til celleprøvning og patogenfri statu...

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Selvom den intranasale rute for lægemiddelafgivelse er blevet udforsket bredt for små molekyler, nanomedicin og proteinforbindelser ens 18 , er anvendelsen af ​​terapeutiske stamceller til intranasal hjernetumor-målretning meget ny i spektret af hjernetumorterapeutik under udvikling 2 , 3 , 4 . Der er involveret egentlige kompleksiteter med hensyn til betegnelsen af ​​stamceller i næsehulen, ...

Lav toksicitet, lav immunogenicitet og indre hjernetumor tropisme af humane stamceller er attraktive træk ved levering af terapeutiske vehikler 1 . Nye stamceller-baserede terapier til maligne hjernetumorer er lovende innovationer udviklet i de senere år, og den intranasale tilpasning af denne terapeutiske strategi er et spring mod klinisk oversættelse, idet ikke-invasiv og gentagen administration dramatisk kan reducere barrieren for patientapplikationer og Kan være tilpasningsberettigede til outpatienttjenester uden generel anæstesi eller langvarig patientbehandling i forbindelse med invasive kirurgiske procedurer 1 , 2 , 3 , 4 . Vi og andre har ført til den intranasale vej for stamcelleudlevering til hjernetumorer og har lagt jorden til at arbejde for nogle af de grundlæggende principperAf translationel forskning ved anvendelse af muse xenograft-modeller 2 , 3 , 4 samt undersøgelse af migration af stamceller in vivo via magnetisk resonansbilleddannelse (MRI) reagensbærere 2 . Gennem disse pilotundersøgelser har vi samlet stor erfaring og fået indsigt i, hvordan man bedst kan opbygge en robust præklinisk evalueringsstrategi ved brug af veletablerede patenterede xenograft (PDX) musemodeller af malignt gliom, samtidig med at undersøgelsesresolutionen opretholdes for at undersøge Nyanserede mekaniske detaljer af de sofistikerede biologiske fænomener af den intranasale hjerneindtrængning af terapeutiske stamceller leveret til næsehulen. Her beskriver vi principperne for en standardiseret driftsprotokol for at demonstrere den aktuelle tilstand af eksperimentelle undersøgelser ved anvendelse af en veletableret human neural stamcellelinje HB1.F3.CD 5 <sup&gt;, 6 , 7 , 8 , som let kan modificeres til at tilpasse sig specifikke tumormodeller eller strategier ved anvendelse af humane stamceller som terapeutiske bærere.

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			<td height="21" style="height:21px;width:236px;">Stereotaxic frame</td>
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			<td height="21" style="height:21px;width:236px;">Hypoxic Cell Culture Incubator</td>
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			<td style="width:119px;">VIOS 160i</td>
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			<td height="21" style="height:21px;width:236px;">Cell culture supplies (Plastics)</td>
			<td style="width:221px;">ThermoFisher Scientific</td>
			<td style="width:119px;">Varies</td>
			<td>Replaceable with any source</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:236px;">Legend Micro 21R Refrigerated Microcentrifuge</td>
			<td style="width:221px;">ThermoFisher Scientific</td>
			<td style="width:119px;">75002490</td>
			<td>Replaceable with any source</td>
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			<td height="21" style="height:21px;width:236px;">Bench centrifuge Sorvall ST16R&#160;</td>
			<td style="width:221px;">ThermoFisher Scientific</td>
			<td style="width:119px;">75004240</td>
			<td>Replaceable with any source</td>
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			<td height="22" style="height:23px;width:236px;">Micro syringe 702N 25&#181;l (22S/2&#34;/2)</td>
			<td style="width:221px;">Hamilton Company</td>
			<td style="width:119px;">80400</td>
			<td>Flat tip</td>
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			<td height="43" style="height:43px;width:236px;">Sample Tray for Irradiator</td>
			<td style="width:221px;">Best Theratronics</td>
			<td style="width:119px;">A13826</td>
			<td style="width:199px;">To set up mice protection with lead shield</td>
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			<td height="21" style="height:21px;width:236px;">Leica DMi8 Microscope</td>
			<td style="width:221px;">Leica Microsystem</td>
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			<td>Custom setup</td>
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			<td height="21" style="height:21px;width:236px;">Leica CM1860 UV cryostat</td>
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			<td>Custom setup</td>
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			<td height="21" style="height:21px;width:236px;">Exel International Insulin Syringe</td>
			<td style="width:221px;">ThermoFisher Scientific</td>
			<td style="width:119px;">14-841-31</td>
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			<td height="21" style="height:21px;width:236px;">Corning Phosphate Buffer Saline</td>
			<td style="width:221px;">Corning Cellgro/ThermoFisher</td>
			<td style="width:119px;">21-031-CV</td>
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			<td height="20" style="height:21px;width:236px;">Dulbecco&#39;s Modified Eagle Medium&#160;</td>
			<td style="width:221px;">Corning Cellgro/ThermoFisher</td>
			<td style="width:119px;">11965-084</td>
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			<td height="21" style="height:21px;width:236px;">Trypsin 0.05%</td>
			<td style="width:221px;">Corning Cellgro/ThermoFisher</td>
			<td style="width:119px;">25300054</td>
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			<td height="21" style="height:21px;width:236px;">Hyaluronidase from bovine testes</td>
			<td style="width:221px;">MilliporeSigma</td>
			<td style="width:119px;">H3506</td>
			<td></td>
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intranasal delivery of therapeutic stem cells to glioblastoma in a mouse model

Den egentlige tropisme mod hjernens maligniteter gør stamceller som lovende bærere af terapeutiske midler mod maligne tumorer. Levering af terapeutiske stamceller via den intranasale vej er en nyligt opdaget alternativ strategi med stærkt potentiale for klinisk oversættelse på grund af dets ikke-invasive karakter sammenlignet med intrakraniel implantation eller levering via systemiske veje. Manglen på blodhjernebarriere styrker yderligere det terapeutiske potentiale hos stamceller, der gennemgår intranasal hjerneindgang. Denne artikel opsummerer de væsentlige teknikker, der anvendes i vores undersøgelser, og beskriver de grundlæggende principper for intranasal strategi for stamcelletilførsel ved hjælp af en musemodel af intrakraniale gliomentraler. Vi demonstrerer de optimerede procedurer, der genererer konsistente og reproducerbare resultater med specifikke forudbestemte eksperimentelle parametre og tilbyder retningslinjer for strømlinet arbejdsstrøm, der sikrer effektiv eksekvering og pålidelig eksperimentNtal udfald. Artiklen er designet til at fungere som en basislinje til yderligere eksperimentel tilpasning baseret på hypotese, stamceller eller tumorspecifikationer.

Både hypoxisk forbehandling ( Figur 4A ) 4 og CXCR4-overekspression ( figur 4B og 4C ) 4 opregner signifikant cellemembran-tilstedeværelsen af ​​CXCR4-receptorer som demonstreret ved flowcytometri. Tumor tropismen demonstreret af NSC&#39;er (blå pile), er vist i tumorvæv histologi (rød cirkel). Tilstedeværelsen af ​​MPIO-mærkede ( Figur 4D</s...

Watch this Scientific Journal Video about Intranasal Delivery of Therapeutic Stem Cells to Glioblastoma in a Mouse Model at JoVE.com

Intranasal Delivery of Therapeutic Stem Cells to Glioblastoma in a Mouse Model

Stamceller er lovende terapeutiske bærere til at behandle hjernetumorer på grund af deres indre tumor tropisme. Ikke-invasiv intranasal stamcelleudlevering omgår blodhjernebarrieren og viser stærkt potentiale for klinisk oversættelse. Denne artikel opsummerer de grundlæggende principper for intranasal stamcelle levering i en musemodel af gliom.

Intranasal levering af terapeutiske stamceller til glioblastom i en musemodel

The intrinsic tropism towards brain malignancies renders stem cells as promising carriers of therapeutic agents against malignant tumors. The delivery of ...

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Medicine

A Mouse Model of in Utero Transplantation

A Mouse Model of in Utero Transplantation

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus.  For example, in ...

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Stereotactic Intracranial Implantation and In vivo Bioluminescent Imaging of Tumor Xenografts in a Mouse Model System of Glioblastoma Multiforme

Stereotaktisk Intrakraniel Implantation og<em&gt; In vivo</em&gt; Bioluminescent Imaging af tumorvæv implanteret i en musemodel System af glioblastoma multiforme

Glioblastoma multiforme (GBM) is a high-grade primary brain cancer with a median survival of only 14.6 months in humans despite standard tri-modality ...

Intranasal Administration of CNS Therapeutics to Awake Mice

Intranasal administration af CNS Therapeutics at vågne mus

Intranasal administration is a method of delivering  therapeutic agents to the central nervous system (CNS). It is  non-invasive and allows large ...

High Throughput Characterization of Adult Stem Cells Engineered for Delivery of Therapeutic Factors for Neuroprotective Strategies

Højt gennemløb Karakterisering af voksne stamceller udviklet til levering af terapeutiske faktorer for Neurobeskyttende Strategies

Mesenchymal stem cells (MSCs) derived from bone marrow are a powerful cellular resource and have been used in numerous studies as potential candidates to ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Studere i bugspytkirtlen Kræft stamceller Karakteristik for udvikling af nye behandlingsstrategier

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

A Neonatal Mouse Spinal Cord Compression Injury Model

En Neonatal Mouse rygmarvskompression Skade Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Vein indskydning Model: En passende model til at studere Bypass Graft Åbenhed

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Image-Guided Resection of Glioblastoma and Intracranial Implantation of Therapeutic Stem Cell-seeded Scaffolds

Image-vejledt resektion af glioblastom og intrakraniel Implantation af terapeutisk stamceller-seedede stilladser

Glioblastoma (GBM), the most common and aggressive primary brain cancer, carries a life expectancy of 12-15 months. The short life expectancy is due in ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

Isometrisk kontraktilitet måling af musen mesenterica arterie ved hjælp af Wire blodkar

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery

Nye metoder for Intranasal Administration Under Inhalation anæstesi til at evaluere næse-til-hjerne Drug Delivery

Intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that circumvents the blood-brain ...