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Xian, China

Zhao D.-Q.,Xian 710032 | Zhao D.-Q.,PLA Fourth Military Medical University | Li Q.,PLA Fourth Military Medical University | Ma Y.,PLA Fourth Military Medical University | And 4 more authors.
Journal of Clinical Rehabilitative Tissue Engineering Research | Year: 2010

BACKGROUND: Tissue engineering has been investigated as a potential means for generation of replacement cartilage. But the ideal seed cell which is still a problem has to be further solved. OBJECTIVE: To observe the solation and cultivation of adipose-derived stromal cells (ADSCs), and to investigate its chondrogenic differentiation ability in vitro. METHODS: Subcutaneous fatty tissue from New Zealand rabbits were digested with collagenase. The obtained ADSCs were cultured in the chondrogenic induced medium. Incubation and passage of ADSCs were observed under inverted microscope. The feasibility of ADSCs cartilage induction was detected by immunohistochemistry, Alcian Blue stain, and picrosirius red stain. Proliferation of ADSCs was observed by MTT methods. RESULTS AND CONCLUSION: ADSCs could be isolated and cultured in vitro. After cultured in chondrogenic medium, the cells differentiated toward cartilage cells and secreted the specific cartilaginous matrices acid mucopolysaccharide and type II collagen. Primary cultured ADSCs were spindle-shaped, with oval nucleus. At day 3, ADSCs were in the exponential growth phase, and the population doubling time was about 55 hours. At day 4, cell growth was stable. At day 9, the cells were observed aging symptoms. Cartilage-induced cell proliferation rate increased significantly, while with the incubation time prolonging, cell volume increased significantly, from long fusiform shape into large, multi-polygonal pseudopodia, furthermore nuclei were also larger, and nucleolus was clear. At day 2 after incubation, ADSCs were in the exponential growth phase, and the population doubling time was about 30 hours. At days 8, the cell growth was stable. After the 15th generation, cell proliferation slowed down and cells were observed signs of aging. This suggested that ADSCs differentiated into chondrocytes under a certain degree; therefore, ADSCs could be served as seed cell for cartilaginous tissue engineering.

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