Study on the biological characteristics of adipose derived stem cell of adult abdomen

Abstract

Objective To isolatie and cultivate human adipose derived stem cell(ASC)from adult abdomen in vitro. To observe and analyse the morphological features of ASC and to test cell surface markers and secretion of cytokines. Methods ASC was isolated from adult abdomen by collagenase digestion method, after that primary culture and subculture were performed. Cell morphology was observed, growth curve was drawn and cell proliferation was evaluated. Ultrastructural morphology of ASC was observed by scanning electron microscopy. Flow cytometry was used to detect the expressions of CD29, CD34, D49d, CD105 and CD106 on the fourth passage of ASC culture. After 1, 3, 6, 24, 72 h of culture, enzyme linked immunosorbant assay (ELISA)method was used to detect contents of transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor (VEGF), basic fibroblast growth factor(bFGF), hepatocyte growth factor(HGF), stromal cell-derived factor 1(SDF1) and interleukin-8 (IL-8) from supernatant of ASC culture. Data were processed with one-way analysis of variance. Results The culture of ASC was successfully isolated in vitro. ASC was stably cultured in a stable passage in vitro, in flat fusiform, fibroblast-like cells shape. ASC morphology was uniform and the growth curve was S-type. Under the scanning electron microscope, spindle shaped cells and rich microvilli on the cell surface were showed. The phenotypes of the fourth passage ASC were positive for CD29, CD49d, CD105, which the rates of expression approximated at 99.6%, 44.5% and 99.5% respectively, while negative for CD34 and CD105, which the rates of expression approximated at 0.6% and 0.7% respectively. TGF-β1, VEGF, bFGF, HGF, SDF1 and IL-8 were secreted, the contents of these cytokines were (56.51±3.61), (129.54±4.08), (39.02±2.64), (73.46±11.75), (396.03±21.31), (149.37±5.48) pg/mL at 1 h after subculture, respectively. And the cytokine contentration increased with culture time, there were significant differences among these (F=47.32, 805.55, 254.89, 69.84, 197.73, 347.52; with P values below 0.05). Conclusions ASC is readily available, grown rapidly and stably with a stable culture and passage. ASC certainly possesses paracrine function and expresses the stem characteristic markers. Key words: Stem cells; Abdomen; Cells, cultured; Cellular structures; Biological markers; Cytokines