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 è consulente di EndGenitor Technologies, Inc. e membro del consiglio di Rimedion Technologies, Inc.

Reagenti e soluzioni EMG-2 supporti (Lonza, N. di cat. Cc-3162 contenente EBM-2 medio basale e EGM-2 Supplementi kit SingleQuot, e fattori di crescita) EBM-2 (Lonza, Cat.. No. cc-3156) integrato con gli integratori intero kit SingleQuot e fattori di crescita (Lonza, Cat.. No. cc-4176), 10% (v / v) di siero fetale bovino (FBS) e 1% (v / v) penicillina (10.000 U / ml) / streptomicina (10.000 pg / ml) / amfotericina (25 pg / ml). Memorizza fino a 1 mese a 4 ° C. Consigl...

<ol>
	<li>Carmeliet, P., Jain, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+and+clinical+applications+of+angiogenesis.">Molecular mechanisms and clinical applications of angiogenesis.</a> Nature. 473, 298-307 (2011).</li><li>Asahara, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+putative+progenitor+endothelial+cells+for+angiogenesis.">Isolation of putative progenitor endothelial cells for angiogenesis.</a> Science. 275, 964-967 (1997).</li><li>Urbich, C., Dimmeler, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+progenitor+cells:+characterization+and+role+in+vascular+biology.">Endothelial progenitor cells: characterization and role in vascular biology.</a> Circ. Res. 95, 343-353 (2004).</li><li>Critser, P. J., Voytik-Harbin, S. L., Yoder, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolating+and+defining+cells+to+engineer+human+blood+vessels.">Isolating and defining cells to engineer human blood vessels.</a> Cell. Prolif. 44, 15-21 (2011).</li><li>Matthias, M., David, N., Josef, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+bench+to+bedside:+what+physicians+need+to+know+about+endothelial+progenitor+cells.">From bench to bedside: what physicians need to know about endothelial progenitor cells.</a> Am. J. Med. 124, 489-4897 (2011).</li><li>Ingram, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vessel+wall-derived+endothelial+cells+rapidly+proliferate+because+they+contain+a+complete+hierarchy+of+endothelial+progenitor+cells.">Vessel wall-derived endothelial cells rapidly proliferate because they contain a complete hierarchy of endothelial progenitor cells.</a> Blood. 105, 2783-276 (2005).</li><li>Ingram, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+a+novel+hierarchy+of+endothelial+progenitor+cells+using+human+peripheral+and+umbilical+cord+blood.">Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood.</a> Blood. 104, 2752-2760 (2004).</li><li>Yoder, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Redefining+endothelial+progenitor+cells+via+clonal+analysis+and+hematopoietic+stem/progenitor+cell+principals.">Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals.</a> Blood. 109, 1801-1809 (2007).</li><li>Reinisch, A., Strunk, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+Animal+Serum+Free+Expansion+of+Human+Umbilical+Cord+Derived+Mesenchymal+Stromal+Cells+(MSCs)+and+Endothelial+Colony+Forming+Progenitor+Cells+(ECFCs.">Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells (MSCs) and Endothelial Colony Forming Progenitor Cells (ECFCs.</a> J. Vis. Exp. (32), e1525 (2009).</li><li>Au, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+in+vivo+potential+of+endothelial+progenitor+cells+from+human+umbilical+cord+blood+and+adult+peripheral+blood+to+form+functional+long-lasting+vessels.">Differential in vivo potential of endothelial progenitor cells from human umbilical cord blood and adult peripheral blood to form functional long-lasting vessels.</a> Blood. 111, 1302-135 (2008).</li><li>Critser, P. J., Kreger, S. T., Voytik-Harbin, S. L., Yoder, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collagen+matrix+physical+properties+modulate+endothelial+colony+forming+cell-derived+vessels+in+vivo.">Collagen matrix physical properties modulate endothelial colony forming cell-derived vessels in vivo.</a> Microvasc. Res. 80, 23-30 (2010).</li><li>Melero-Martin, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+robust+and+functional+vascular+networks+in+vivo+with+human+adult+and+cord+blood-derived+progenitor+cells.">Engineering robust and functional vascular networks in vivo with human adult and cord blood-derived progenitor cells.</a> Circ. Res. 103, 194-202 (2008).</li><li>Melero-Martin, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+vasculogenic+potential+of+human+blood-derived+endothelial+progenitor+cells.">In vivo vasculogenic potential of human blood-derived endothelial progenitor cells.</a> Blood. 109, 4761-4768 (2007).</li><li>Hofmann, N. A., Reinisch, A., Strunk, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+Large+Scale+Expansion+of+Adult+Human+Endothelial+Colony+Forming+Progenitor+Cells.">Isolation and Large Scale Expansion of Adult Human Endothelial Colony Forming Progenitor Cells.</a> J. Vis. Exp. (32), e1524 (2009).</li><li>Lin, Y., Weisdorf, D. J., Solovey, A., Hebbel, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Origins+of+circulating+endothelial+cells+and+endothelial+outgrowth+from+blood.">Origins of circulating endothelial cells and endothelial outgrowth from blood.</a> J. Clin. Invest. 105, 71-77 (2000).</li><li>Witting, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+Large+Volume+Lentiviral+Vector+Production+Using+Flow+Electroporation.">Efficient Large Volume Lentiviral Vector Production Using Flow Electroporation.</a> Hum. Gene. Ther. , (2011).</li><li>Yoon, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+neovascularization+by+mixed+transplantation+of+early+endothelial+progenitor+cells+and+late+outgrowth+endothelial+cells:+the+role+of+angiogenic+cytokines+and+matrix+metalloproteinases.">Synergistic neovascularization by mixed transplantation of early endothelial progenitor cells and late outgrowth endothelial cells: the role of angiogenic cytokines and matrix metalloproteinases.</a> Circulation. , 112-1618 (2005).</li><li>Dubois, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+effects+of+progenitor+cell+populations+on+left+ventricular+remodeling+and+myocardial+neovascularization+after+myocardial+infarction.">Differential effects of progenitor cell populations on left ventricular remodeling and myocardial neovascularization after myocardial infarction.</a> J. Am. Coll. Cardiol. 55, 2232-2243 (2010).</li><li>Medina, R. J., O'Neill, C. L., Humphreys, M. W., Gardiner, T. A., Stitt, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Outgrowth+endothelial+cells:+characterization+and+their+potential+for+reversing+ischemic+retinopathy.">Outgrowth endothelial cells: characterization and their potential for reversing ischemic retinopathy.</a> Invest. Ophthalmol. Vis. Sci. 51, 5906-5913 (2010).</li></ol>

Caratterizzazione fenotipica e funzionale delle cellule progenitrici endoteliali putativi è importante identificare le ECFCs buona fede che sono in grado di clonale serialmente re-placcatura in coltura e dare origine a durevoli e funzionali vasi sanguigni impiantabili in vivo. Sangue del cordone ombelicale umano si arricchisce di ECFCs e la concentrazione di queste cellule circolanti diminuisce con l&#39;invecchiamento o malattia 10. Recenti studi suggeriscono che ECFC può giocare un ruolo...

<table border="1"><tbody><tr><td> Nome del reattivo </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Iniezione di sodio eparina, USP </td><td> APP Pharmaceuticals </td><td> 504031 </td></tr><tr><td> Ficoll-Pague </td><td> Amersham Biosciences </td><td> 17-1440-03 </td></tr><tr><td> Cannula di miscelazione </td><td> Maersk Medical </td><td> 500.11.012 </td></tr><tr><td> EGM-2 </td><td> Lonza </td><td> CC-3162 </td></tr><tr><td> Definito FBS </td><td> Hyclone </td><td> SH30070.03 </td></tr><tr><td> Triplo express </td><td> Gibco </td><td> 12605 </td></tr><tr><td> Rat collagene di tipo I </td><td> BD Biosciences </td><td> 354236 </td></tr><tr><td> Matrigel </td><td> BD Biosciences </td><td> 356234 </td></tr><tr><td> FcR Block </td><td> Miltenyi Biotech </td><td> 130-059-901 </td></tr><tr><td> hCD31, FITC coniugazioneTed </td><td> BD Pharmingen </td><td> 555445 </td></tr><tr><td> hCD45, coniugato FITC </td><td> BD Pharmingen </td><td> 555482 </td></tr><tr><td> hCD14, coniugato FITC </td><td> BD Pharmingen </td><td> 555397 </td></tr><tr><td> hCD144, PE coniugato </td><td> eBioscience </td><td> 12-1449-80 </td></tr><tr><td> hCD146, PE coniugato </td><td> BD Pharmingen </td><td> 550315 </td></tr><tr><td> hCD105, PE coniugato </td><td> Invitrogen </td><td> MHCD10504 </td></tr><tr><td> Ms 1 IgG, anticorpi k, coniugato FITC </td><td> BD Pharmingen </td><td> 555748 </td></tr><tr><td> Ms 1 IgG, anticorpi k, coniugato con PE </td><td> BD Pharmingen </td><td> 559320 </td></tr><tr><td> Ms IgG 2a, anticorpi k, coniugato FITC </td><td> BD Pharmingen </td><td> 555573 </td></tr><tr><td> Anti-CD31 umano </td><td> Dako </td><td> clone JC70 / A </td></tr><tr><td> Anti-mouse CD31 </td><td> BD Pharmingen </td><td> 553370 </td></tr><tr><td> 0,22-um vuoto sistema di filtrazione </td><td> Millipore </td><td> SCGPU05RE </td></tr><tr><td> Acido acetico glaciale, 17.4N </td><td> Pescatore </td><td> A38-500 </td></tr><tr><td> Antibiotico-antimicotica </td><td> Invitrogen </td><td> 15240-062 </td></tr><tr><td> Siero bovino fetale (FBS) </td><td> Hyclone </td><td> SH30070.03 </td></tr><tr><td> IHC zinco Fixative </td><td> BD Biosciences </td><td> 550523 </td></tr><tr><td> Sytox verde reagente </td><td> Invitrogen </td><td> S33025 </td></tr><tr><td> Cilindri clonazione, sterili </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

Viste di vecchia data della formazione di nuovi vasi sanguigni attraverso l&#39;angiogenesi, vasculogenesi e arteriogenesi sono stati recentemente rivisti 1. La presenza in circolo di cellule progenitrici endoteliali (EPC) sono stati identificati nel sangue periferico umano adulto da Asahara et al. Nel 1997 2 portando un&#39;infusione di nuove ipotesi e strategie per la rigenerazione e la riparazione vascolare. EPC sono componenti rari, ma di normale circolazione del sangue, che ospita i siti di formazione dei vasi sanguigni o di rimodellamento vascolare, e facilitare sia vasculogenesi post-natale, l&#39;angiogenesi, o arteriogenesi in gran parte attraverso la stimolazione paracrina della parete del vaso esistente derivato cellule 3. Nessun marcatore specifico per identificare un EPC è stato identificato, e al momento lo stato del campo è quello di capire che tipi di cellule tra cui numerosi proangiogenica staminali e cellule progenitrici ematopoietiche, cellule circolanti angiogenici, Tie2 + monociti, cel progenitore mieloidels, tumore macrofagi associati, e M2 macrofagi attivati ​​partecipare stimolare il processo angiogenico in una varietà di sistemi preclinici modelli animali e in soggetti umani in numerosi stati di malattia 4, 5. Le cellule endoteliali che formano colonie (ECFCs) sono rare cellule vitali endoteliali circolanti caratterizzate da una forte potenziale proliferativo clonale, colonia secondaria e terziaria capacità di formazione su replating, e la capacità di formare intrinseca in vasi vivo dopo il trapianto in topi immunodeficienti 6-8. Mentre ECFCs sono state isolate con successo dal sangue periferico di soggetti adulti sani, sangue del cordone ombelicale (CB) di neonati sani e parete del serbatoio di numerosi vasi arteriosi e venosi umane 6-9, CB possiede la più alta frequenza di ECFCs 7 che mostra il più robusto potenziale proliferativo clonale e la forma dei vasi sanguigni durevoli e funzionali in vivo 8, 10-13. Sebbene la derivazione diECFC adulto da sangue periferico è stato presentato 14, 15, qui si descrivono le metodologie per la derivazione, clonazione, l&#39;espansione, e in vitro e in vivo caratterizzazione del ECFCs dal ombelicale umano 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

Le cellule endoteliali che formano colonie (ECFCs) sono cellule endoteliali circolanti con una robusta potenziale proliferativo clonale che mostrano intrinseca<em&gt; In vivo</emNave&gt; capacità di formazione. Caratterizzazione fenotipica e funzionale delle cellule endoteliali escrescenza derivate da CB sono importanti per identificare ed isolare<em&gt; Bona fide</em&gt; ECFCs per il potenziale applicazione clinica nella riparazione di tessuti danneggiati.

Caratterizzazione fenotipica e funzionale delle cellule endoteliali Colony Forming derivato dal sangue del cordone ombelicale umano

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

Research

JoVE Journal

Biology

Isolation of Human Umbilical Vein Endothelial Cells (HUVEC)

Isolation of Human Umbilical Vein Endothelial Cells HUVEC

Isolamento di cellule umane endoteliali della vena ombelicale (HUVEC)

Angiogenesis is a complex multi-step process, where in response to angiogenic stimuli, new vessels are created from the existing vasculature. These steps ...

Ricerca

Biologia

Isolation and Large Scale Expansion of Adult Human Endothelial Colony Forming Progenitor Cells

Isolamento e di espansione su vasta scala di umano adulto cellule progenitrici endoteliali che formano colonie

This paper introduces a novel recovery strategy for endothelial colony forming progenitor cells (ECFCs) from heparinized but otherwise unmanipulated adult ...

Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells (MSCs) and Endothelial Colony Forming Progenitor Cells (ECFCs)

Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells MSCs and Endothelial Colony Forming Progenitor Cells ECFCs

Isolamento e di espansione degli animali siero gratuito di cordone ombelicale umano derivato cellule stromali mesenchimali (MSC) e cellule endoteliali progenitrici formanti colonia (ECFCs)

The umbilical cord is a rich source for progenitor cells with high proliferative potential including mesenchymal stromal cells (also termed mesenchymal ...

Isolation and Enrichment of Rat Mesenchymal Stem Cells (MSCs) and Separation of Single-colony Derived MSCs

Isolation and Enrichment of Rat Mesenchymal Stem Cells MSCs and Separation of Single-colony Derived MSCs

L&#39;isolamento e l&#39;arricchimento delle cellule staminali mesenchimali di ratto (MSC) e la separazione dei Single-colonia di cellule staminali mesenchimali derivate

MSCs are a population of adult stem cells that is a promising source for therapeutic applications. These cells can be isolated from the bone marrow and ...

Isolation of Human Umbilical Arterial Smooth Muscle Cells (HUASMC)

Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC

Isolamento di cellule umane ombelicale muscolatura liscia arteriosa (HUASMC)

The human umbilical cord (UC) is a biological sample that can be easily obtained just after birth. This biological sample is, most of the time, discarded ...

Isolation and Culture of Pulmonary Endothelial Cells from Neonatal Mice

Isolamento e la coltura di cellule endoteliali polmonari da topi neonati

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells ...

A Quantitative Assay for Insulin-expressing Colony-forming Progenitors

Un dosaggio quantitativo di insulina che esprimono formante colonia Progenitori

The field of pancreatic stem and progenitor cell biology has been hampered by a lack of in vitro functional and quantitative assays that allow for the ...

Manual Isolation of Adipose-derived Stem Cells from Human Lipoaspirates

Isolamento Manuale di cellule staminali derivate da tessuto adiposo da lipoaspirato umani

In 2001, researchers at the University of  California, Los Angeles, described the isolation of a new population of adult  stem cells from liposuctioned ...

Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

Isolamento dei vasi sanguigni derivati ​​da Multipotent precursori del muscolo scheletrico umano

Since the discovery of mesenchymal stem/stromal cells (MSCs), the native identity and localization&#160;of MSCs have been obscured by their retrospective ...

Two-photon Imaging of Intracellular Ca2+ Handling and Nitric Oxide Production in Endothelial and Smooth Muscle Cells of an Isolated Rat Aorta

Two-photon Imaging of Intracellular Ca2+ Handling and Nitric Oxide Production in Endothelial and Smooth Muscle Cells of an Isolated Rat Aorta

A due fotoni Imaging di Intracellulare Ca<sup&gt; 2+</sup&gt; Manipolazione e l&#39;ossido nitrico Produzione in endoteliale e della muscolatura liscia cellule di un isolato Rat Aorta

Calcium is a very important regulator of many physiological processes in vascular tissues. Most endothelial and smooth muscle functions highly depend on ...