脂肪间充质干细胞与自体髓核细胞共培养中相互作用分子机制初步研究.pdfVIP

华 中 科 技 大 学 硕 士 学 位 论 文 In Vitro Study on Interaction Between Adipose Tissue-derived Mesenchymal Stem Cells and Autologous Nucleus Pulposus Cells Through Paracrine Stimulation Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology Candidate: Xu Zhi-Gang Supervisor: Prof. Fang Huang Abstract Objective Co-culture adipose tissue-derived mesenchymal stem cells (AMSCs )and autologous nucleus pulposus(NP) cells without direct contacting under hypoxia. To investigate the interaction between NP cells and AMSCs through paracrine stimulation. Methods ①Rat AMSCs were isolated from the subcutaneous inguinal region only, purified by limited dilution. ②AMSCs of the third passage were analyzed by Fluorescence Activated Cell Sorter (FASC) to detect the cell surface markers. ③Co-culture AMSCs and degenerative NP cells without direct contacting in different ratios under hypoxia. ④Two weeks later, NP cells proliferation and RNA expression of selected genes were then evaluated after co-cultured; immunohistochemical analysis and RNA expression were used to detect AMSCs after induced. Results ①Proliferation of NP cells significantly rose even after cocultured with a few AMSCs. The biologic and metabolic viability of NP cells were upregulated, the results of RT-PCR manifested that there was significant difference in gene expression of aggrecan 4 华 中 科 技 大 学 硕 士 学 位 论 文 and collagen II among two groups (P0.01). Immunohistochemical analysis showed that AMSCs started to express collagen II, the results of RT-PCR demonstrat