The isolation, subculture and identification of human adipose derived endothelial progenitor cells / 中华整形外科杂志
Chinese Journal of Plastic Surgery
;
(6): 62-65, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-297095
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the methods of isolating and identifying human adipose derived EPCs.</p><p><b>METHODS</b>The cells obtained from human lipoaspirates were plated on culture dishes coated with human fibronectin and were cultured in DMEM containing 2% FBS. Cells of passage 2 cultured in EGM-2 (2% FBS) served as the induced cells (experimental group), with cells cultured in DMEM (2% FBS) as the non-induced cells (control group) . Immunofluorescence was used to detect the expression of cell markers, including CD34, vWF and PECAM-1. FACS (fluorescence activated cell sorter) was used to quantitatively analyze the expression rate of cell markers (CD34, CD45, CD133 and PECAM-1). Fluorescence microscope was used to observe the function of taking up DiI-ac-LDL by the induced cells. To determine the ability of forming capillary-like structure in three-dimensional matrices, the induced cells were also cultured in methylcellulose.</p><p><b>RESULTS</b>The induced cells of passage 2 exhibited cobblestone morphology, similar to that of the endothelial cells. In contrast, these morphological changes were not observed in non-induced cells. Immunofluorescence detected expression of vWF, PECAM-1 in induced cells and CD34 in non-induced cells. FACS analysis showed (67.41 +/- 13.35)% of the induced cells expressed PECAM-1 and (6.73 +/- 2.21)% of the non-induced cells expressed PECAM-1 (P < 0.01), while (72.39 +/- 13.45)% of the non-induced cells expressed CD34 and (16.06 +/- 3.86)% of the induced cells expressed CD34 (P < 0.01). Fluorescence microscopy observed the induced cells took up low-density lipoprotein (LDL). The formation of "branch-like" structure confirmed their functional activity.</p><p><b>CONCLUSION</b>EPCs derived from human adipose may serve as another source of seeding cells for vascular tissue engineering.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Stem Cells
/
Cell Count
/
Cell Differentiation
/
Cell Movement
/
Cells, Cultured
/
Adipocytes
/
Cell Culture Techniques
/
Cell Biology
/
Endothelial Cells
/
Cell Proliferation
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Plastic Surgery
Year:
2007
Type:
Article
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