Name: phenotypic and functional characterization of endothelial colony forming cells derived from human umbilical cord blood
Uploaded: 2012-04-13T13:00:00
Description: Watch this Scientific Journal Video about Phenotypic and Functional Characterization of Endothelial Colony Forming Cells Derived from Human Umbilical Cord Blood at JoVE.com

Dr Yoder är konsult EndGenitor Technologies, Inc. och ledamot i styrelsen för Rimedion Technologies, Inc.

Reagens och lösningar EMG-2 media (Lonza, Cat. Nr cc-3162 innehåller EBM-2 basalmedium och EGM-2 SingleQuot Kosttillskott kit, &amp; tillväxtfaktorer) EBM-2 (Lonza, kat nr. Cc-3156) kompletterat med hela tillskott SingleQuot kit och tillväxtfaktorer (Lonza, Kat nr. Cc-4176), 10% (volym / volym) fetalt bovint serum (FBS) och 1% (volym / volym) penicillin (10.000 U / ml) / streptomycin (10.000 xg / ml) / amfotericin (25 pg / ml). Lagra upp till 1 månad vid 4 ° C. ...

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Fenotypisk och funktionell karakterisering av förmodade endotelceller stamceller är det viktigt att identifiera de seriösa ECFCs som kan klonalt och seriellt Re-plating i kultur och ge upphov till långsiktiga och funktionella implanterbara blodkärl in vivo. Humant navelsträngsblod är berikad med ECFCs och koncentrationen av dessa cirkulerande celler minskar med åldrandet eller sjukdom 10. Nyligen genomförda studier tyder på att ECFC kan spela viktiga roller i vaskulär reparation e...

<table border="1"><tbody><tr><td> Namnet på reagens </td><td> Företaget </td><td> Katalognummer </td></tr><tr><td> Heparinnatrium, USP </td><td> APP Läkemedel </td><td> 504.031 </td></tr><tr><td> Ficoll-Pague </td><td> Amersham Biosciences </td><td> 17-1440-03 </td></tr><tr><td> Blandning kanyl </td><td> Maersk Medicinsk </td><td> 500.11.012 </td></tr><tr><td> EGM-2 </td><td> Lonza </td><td> CC-3162 </td></tr><tr><td> Definieras FBS </td><td> Hyclone </td><td> SH30070.03 </td></tr><tr><td> TrypLE uttrycker </td><td> Gibco </td><td> 12.605 </td></tr><tr><td> Råtta typ I-kollagen </td><td> BD Biosciences </td><td> 354.236 </td></tr><tr><td> Matrigel </td><td> BD Biosciences </td><td> 356.234 </td></tr><tr><td> FcR Block </td><td> Miltenyi Biotech </td><td> 130-059-901 </td></tr><tr><td> hCD31, FITC conjugaTed </td><td> BD Pharmingen </td><td> 555.445 </td></tr><tr><td> hCD45, FITC-konjugerad </td><td> BD Pharmingen </td><td> 555.482 </td></tr><tr><td> hCD14, FITC-konjugerad </td><td> BD Pharmingen </td><td> 555.397 </td></tr><tr><td> hCD144, PE-konjugerad </td><td> eBioscience </td><td> 12-1449-80 </td></tr><tr><td> hCD146, PE-konjugerad </td><td> BD Pharmingen </td><td> 550.315 </td></tr><tr><td> hCD105, PE-konjugerad </td><td> Invitrogen </td><td> MHCD10504 </td></tr><tr><td> Ms IgG 1, k antikropp, FITC-konjugerad </td><td> BD Pharmingen </td><td> 555.748 </td></tr><tr><td> Ms IgG 1, k antikropp, PE-konjugerad </td><td> BD Pharmingen </td><td> 559.320 </td></tr><tr><td> Ms IgG2a, k antikropp, FITC-konjugerad </td><td> BD Pharmingen </td><td> 555.573 </td></tr><tr><td> Anti-human CD31 </td><td> Dako </td><td> klon JC70 / A </td></tr><tr><td> Anti-mouse CD31 </td><td> BD Pharmingen </td><td> 553.370 </td></tr><tr><td> 0,22 | im vakuumfiltrering systemet </td><td> Millipore </td><td> SCGPU05RE </td></tr><tr><td> Isättika, 17.4N </td><td> Fisher </td><td> A38-500 </td></tr><tr><td> Antibiotika-Antimykotisk </td><td> Invitrogen </td><td> 15240-062 </td></tr><tr><td> Fetalt bovint serum (FBS) </td><td> Hyclone </td><td> SH30070.03 </td></tr><tr><td> IHC Zink Fixativ </td><td> BD Biosciences </td><td> 550.523 </td></tr><tr><td> Sytox grön reagens </td><td> Invitrogen </td><td> S33025 </td></tr><tr><td> Kloning cylindrar, sterila </td><td> Fisher Scientific </td><td> 07-907-10 </td></tr></tbody></table>

phenotypic and functional characterization of endothelial colony forming cells derived from human umbilical cord blood

Långvariga utsikt över New blodkärl via angiogenes, vaskulogenes och arteriogenes har nyligen omdömet 1. Förekomsten av cirkulerande endotelceller stamceller (EPC) först identifierades i vuxen människa perifert blod genom Asahara et al. 1997 2 väcka infusion av nya hypoteser och strategier för vaskulär regeneration och reparation. EPCs är sällsynta, men normala komponenter i cirkulerande blod som hem till platser för blodkärlsbildning eller vaskulär remodellering, och underlätta antingen postnatal vaskulogenes, angiogenes, eller arteriogenes till stor del via olika typer av stimulering av befintlig kärlväggen härrör celler 3. Ingen specifik markör för att identifiera en EPC har identifierats, och för närvarande tillstånd av fältet är att förstå att många celltyper inklusive proangiogen hematopoetiska stam-och progenitorceller, cirkulerande angiogena celler, Tie2 + monocyter, myeloid progenitor cells, tumörassocierade makrofager, och M2 aktiverade makrofager delta i att stimulera den angiogena processen i en mängd av prekliniska djurmodellsystem och humanpatienter i ett flertal sjukdomstillstånd 4, 5. Endotelceller kolonibildande celler (ECFCs) är sällsynta cirkulerande livskraftiga endotelceller kännetecknas av en stark klonal proliferativ potential, sekundär och tertiär kolonibildande förmågan vid återutstrykning och förmåga att bilda inneboende in vivo kärl vid transplantation till immundefekta möss 6-8. Även ECFCs har framgångsrikt isolerats från perifert blod hos friska vuxna individer, navelsträngsblod (CB) av friska nyfödda barn, och kärlväggen av många mänskliga arteriella och venösa kärl 6-9, har CB den högsta frekvensen av ECFCs 7 som visas den mest robusta klonal proliferativ potential och form hållbara och funktionella blodkärl in vivo 8, 10-13. Medan härledning avECFC från perifert blod från vuxna har presenterats 14, 15, här beskriver metoder för härledning, kloning, expansion, och in vitro såväl som in vivo karakterisering av ECFCs från humana umbilikala CB.

Watch this Scientific Journal Video about Phenotypic and Functional Characterization of Endothelial Colony Forming Cells Derived from Human Umbilical Cord Blood at JoVE.com

Phenotypic and Functional Characterization of Endothelial Colony Forming Cells Derived from Human Umbilical Cord Blood

Endotelceller kolonibildande celler (ECFCs) cirkulerar endotelceller med robust klonal proliferativ potential som visar inneboende<em&gt; In vivo</em&gt; Fartyget en förmåga till bildning. Fenotypisk och funktionell karakterisering av utväxt endotelceller som härrör från CB är viktigt att identifiera och isolera<em&gt; Bona fide</em&gt; ECFCs för potentiella klinisk tillämpning för att reparera skadade vävnader.

Fenotypisk och funktionell karakterisering av Endotelceller kolonibildande celler härledda från humant navelsträngsblod

Longstanding views of new blood vessel formation via angiogenesis, vasculogenesis, and arteriogenesis have been recently reviewed1. The presence of ...

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