Abstract:

BACKGROUND: Adipose-derived stem cells (ADSCs) are easily collected and abundantly cultured, ADSCs can proliferate rapidly when being cultured in vitro, with multi-directional differentiation potential, ADSCs are expected as seed cells for tissue engineering and cell therapy.
OBJECTIVE: To culture rat ADSCs in vitro, and to mark and identify its differentiation.
METHODS: CollagenaseⅠdigestion was used to isolate ADSCs from rat groin fat pads under sterile condition. ADSCs were passaged and amplified by the trypsin digestion. At the third passage, growth curve of the third passage ADMSCs was drawn by cytometry. Cellular surface antigens HCAM, CD106, CD29, CD49D and CD34 were examined using flow cytometry. Giemsa staining, Feridex-labeled and induced to differentiate into osteoblasts and lipocytes. Oil red staining was used to examine lipocyte induction, alizarin red staining wwas used to examine osteoblast induction.
RESULTS AND CONCLUSION: In vitro cultured ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. The third passage ADSCs were positive for CD29, but negative for CD34, HCAM, CD49d and CD106. Curve of growth detection had logarithmic phase and platform phase, the positive expression of Feridex reached 80%. ADMSCs were differentiated into lipocytes and osteoblasts under certain conditions. ADSCs isolated from rat epididymal fat pads can be easily cultured, passaged and amplified, and which can mark alive and differentiate into osteoblasts and adipocytes under certain conditions.