Differentiation of human embryonic stem cells to endothelial cells via improved three-dimension approach / 中国医学科学院学报
Acta Academiae Medicinae Sinicae
;
(6): 539-544, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-284336
ABSTRACT
<p><b>OBJECTIVE</b>To establish an improved three-dimension (3D) and serum-free approach to differentiate human embryonic stem cells (hESCs) into endothelial cells, and detect the endothelial functions of the obtained cells.</p><p><b>METHODS</b>We cultured undifferentiated H9 human embryonic stem cell line in low-adhesion dishes to form embryonic bodies (EBs). After 12 days, EBs were harvested, re-suspended into rat tail collagen type I, and put into the incubator (37℃). After 30 minutes, EGM-2 culture medium was added to the solidified collagen, and the EBs were cultured for another 3 days to form embryonic body-sproutings (EB-sproutings). EB-sproutings were digested with 0.25% collagenase I and 0.56 U/ml Liberase Blendzyme for 20 minutes respectively, and the CD31(+) cells were sorted by FACS. The endothelial functions were tested by Dil-ac-LDL uptake assay and tube formation assay.</p><p><b>RESULTS</b>This approach raised the efficiency of endothelial differentiation to 18%, and also avoided the contamination with animal materials. The obtained hESC-derived endothelial cells (hESC-ECs) had the similar pattern of surface biomarkers as human umbilical vein endothelial cells (HUVECs), and their endothelial functions were confirmed by the uptake of Dil-ac-LDL and the tube formation on Matrigel.</p><p><b>CONCLUSIONS</b>The improved 3D approach can enhance the efficiency of differentiation from hESCs into endothelial cells. Furthermore, serum free differentiation system may be applied in future hESC-based therapies for various ischemic diseases.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Cell Differentiation
/
Cell Line
/
Cell Culture Techniques
/
Culture Media
/
Cell Biology
/
Collagen Type I
/
Endothelial Cells
/
Embryonic Stem Cells
/
Methods
Limits:
Humans
Language:
Chinese
Journal:
Acta Academiae Medicinae Sinicae
Year:
2012
Type:
Article
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