[Study on culture, identification and differentiation of CD133+ endothelial progenitor cells from human umbilical cord blood].
OBJECTIVE
To study the isolation, culture and identification of CD133+ endothelial progenitor cells (EPCs) from human umbilical cord blood in vitro.


METHODS
EPC separation was performed with density gradient centrifugation and MACS separation. Purity of EPCs was determined by flow cytometry. EPC was cultured with EBM-2 to study the cultivate features of EPC. Uptake test of Dil-LDL and FITC-Lectin and immunohistochemistry were performed.


RESULTS
According to flow cytometry, (1.13 +/- 0.10)% of mono-nuclear cells were CD133+ and the purities of CD133+ EPCs were (91.45 +/- 1.04)% on average. CD133+ EPCs became adherent, spindle-shaped and formed cluster during culture. Uptake test of Dil-LDL and FITC-Lectin were positive. (95.83 +/- 1.72)% of CD133+ cells were found positive in both uptake tests. The positive rates of immunostaining of cell markers CD34 and factor VIII were (95.83 +/- 2.23)% and (95.92 +/- 1.43)% after cultured for one week, which showed no significant differences between CD133+ EPCs and human umbilical vein endothelial cells. Capillary structures were formed by CD133+ EPCs after cultured for 4 and 7 d in vitro.


CONCLUSIONS
High purity of CD133+ EPCs can be obtained by MACS separation. CD133+ EPCs can differentiate into mature endothelial cells with the effects of stimulating factors.
Huang
Yong
Y
Department of Vascular Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
Wang
Shen-ming
SM
Wang
Jin-song
JS
Huang
Xue-ling
XL
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000032:analysis* / Q000097:blood; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000276:immunology; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology; D005434:Flow Cytometry; D006023:Glycoproteins / Q000032:analysis* / Q000097:blood; D006801:Humans; D018189:Immunomagnetic Separation; D010455:Peptides / Q000032:analysis* / Q000097:blood; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology
D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides