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 arbeidet ble støttet av NIH R01NS087990 (MSL, IVB).

Alle dyreprosedyrer må godkjennes av Institutional Animal Care and Use Committee (IACUC) eller tilsvarende. Hvis det er noen usikkerhet om de spesifikke prosedyrene beskrevet her, fortsett ikke. Avklare med institusjonens IACUC og et utpekt veterinærmedarbeider. 1. Sikre sterilitet av dyrkede celler og følg prinsippene for aseptiske teknikker <ol><li> Følg standard god laboratorieøvelse i alle cellekulturprosedyrer og IACUC-krav for celleprøving og patogenfri statusbekreftelse før a...

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Selv om den intranasale ruten for legemiddellevering er blitt utbredt for små molekyler, nanomedisiner og proteinforbindelser likt 18 , er anvendelsen av terapeutiske stamceller for intranasal hjernesvulster-måling meget ny i spekteret av hjernetumorbehandlingene under utvikling 2 , 3 , 4 . Det er inneboende kompleksiteter involvert angående betegnelsen av stamceller i neshulen, og molekylære detalje...

Lav toksisitet, lav immunogenicitet og egenartet hjerne-tumor-tropisme av menneskelige stamceller er attraktive egenskaper for levering av terapeutiske kjøretøy 1 . Nye stamceller-baserte terapeutiske midler for ondartede hjernesvulster er lovende innovasjoner utviklet de siste årene, og den intranasale tilpasningen av denne terapeutiske strategien representerer et skritt mot klinisk oversettelse, fordi ikke-invasiv og gjentatt administrasjon kan dramatisk redusere barrieren for pasientapplikasjoner og Kan være tilpasningsdyktig for pasientrelaterte tjenester uten generell bedøvelse eller langvarig pasientstjeneste i forbindelse med invasive kirurgiske prosedyrer 1 , 2 , 3 , 4 . Vi og andre har pioneret den intranasale ruten for stamcelleavlasting til hjernesvulster og har lagt grunnarbeidet for noen av de grunnleggende prinsippeneAv translasjonsforskning ved hjelp av musen xenograft-modeller 2 , 3 , 4 , samt undersøkte migrering av stamceller in vivo via magnetisk resonans imaging (MR) reagensbærere 2 . Gjennom disse pilotundersøkelsene har vi samlet stor erfaring og fått innsikt i hvordan man best kan konstruere en robust preklinisk evalueringsstrategi ved å bruke veletablerte pasient-avledede xenograft (PDX) musemodeller av ondartet gliom, samtidig som undersøkelsesoppløsningen ble holdt for å undersøke Nyanserte mekaniske detaljer om de sofistikerte biologiske fenomenene av den intranasale hjernen oppføring av terapeutiske stamceller levert til nesehulen. Her beskriver vi prinsippene for en standardisert driftsprotokoll for å demonstrere den nåværende tilstanden for eksperimentelle undersøkelser ved å bruke en veletablert human neural stamcellelinje HB1.F3.CD 5 <sup&gt;, 6 , 7 , 8 , som lett kan modifiseres til å tilpasse seg bestemte tumormodeller eller strategier ved bruk av 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>
			<td></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>
		</tr>
		<tr height="21">
			<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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		<tr height="21">
			<td height="21" style="height:21px;width:236px;">Leica DMi8 Microscope</td>
			<td style="width:221px;">Leica Microsystem</td>
			<td style="width:119px;"></td>
			<td>Custom setup</td>
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			<td height="21" style="height:21px;width:236px;">Leica CM1860 UV cryostat</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;">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 tropismen mot maligniteter i hjernen gjør stamceller som lovende bærere av terapeutiske midler mot maligne tumorer. Leveransen av terapeutiske stamceller via den intranasale ruten er en nylig oppdaget alternativ strategi, med sterkt potensial for klinisk oversettelse, på grunn av sin ikke-invasive natur sammenlignet med intrakranial implantasjon eller levering via systemiske veier. Mangelen på blodhjernebarriere styrker det terapeutiske potensialet til stamceller som undergår intranasal hjerneoppføring. Denne artikkelen oppsummerer de grunnleggende teknikkene som brukes i våre studier, og skisserer de grunnleggende prinsippene for intranasal strategi for stamcelleavlasting ved hjelp av en musemodell av intrakraniell gliomentral. Vi demonstrerer de optimaliserte prosedyrene som genererer konsistente og reproducerbare resultater med bestemte forhåndsbestemte eksperimentelle parametere og gir retningslinjer for strømlinjeformet arbeidsflyt som sikrer effektiv utførelse og pålitelig eksperimentNtal utfall. Artikkelen er utformet for å fungere som en basislinje for videre eksperimentell tilpasning basert på hypotese, stamcelletyper eller svittspesifikasjoner.

Både hypoksisk forbehandling ( Figur 4A ) 4 og CXCR4-overekspresjon ( figur 4B og 4C ) 4 oppregner signifikant cellemembran-nærværet av CXCR4-reseptorer som vist ved strømningscytometri. Tumor tropismen demonstrert av NSCs (blå piler), er vist i tumorvevshistologi (rød sirkel). Tilstedeværelsen av MPIO-merkede ( Figur 4D ) stamceller i svuls...

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 å behandle hjernesvulster på grunn av deres egen tumor-tropisme. Ikke-invasiv intranasal stamcelleavlevering omgår blodhjernebarrieren og viser sterkt potensial for klinisk oversettelse. Denne artikkelen oppsummerer de grunnleggende prinsippene for intranasal stamcelle levering i en musemodell av gliom.

Intranasal levering av terapeutiske stamceller til glioblastom i en musemodell

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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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

Stereotactic Intrakraniell Implantasjon og<em&gt; In vivo</em&gt; Bioluminescent Imaging av tumorxenotransplantater i en mus modell System of 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

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

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

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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

Bilde-guidede Resection av glioblastom og intrakranielt implantering av terapeutiske Stem Cell-seeded stillaser

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 ...

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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

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