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目的探索兔骨髓间充质干细胞向软骨组织转化的条件,为组织工程寻找新的工程化软骨。方法用密度梯度离心法和贴壁法分离纯化骨髓间充质干细胞,并传代扩增后,用离心三维培养法在成软骨诱导剂下进行诱导培养,并分别在诱导培养第7、14、21天取出标本并进行组织切片,对标本行免疫组织化学、原位杂交检测。结果骨髓间充质干细胞经7、14、21d的离心三维诱导培养后,培养管出现软骨外观组织块,组织学切片可见软骨陷窝,进行Ⅱ型胶原的免疫组织化学、原位杂交检测均为阳性。结论兔骨髓间充质干细胞在软骨诱导剂和离心管培养条件下可成功生成软骨组织。
Objective To explore the conditions of rabbit bone marrow mesenchymal stem cells into cartilage tissue, looking for new engineered cartilage for tissue engineering. Methods Bone marrow mesenchymal stem cells (MSCs) were isolated and purified by density gradient centrifugation and adherent method. After passaged and expanded, the cells were cultured under the condition of cartilage induction by centrifugation and three-dimensional culture. The cells were cultured for 7, 14, 21 The specimens were removed and the tissue sections were taken. The specimens were examined by immunohistochemistry and in situ hybridization. Results After the bone marrow mesenchymal stem cells were three-dimensionally induced by centrifugation on 7, 14, and 21 days, cartilage appearance tissue mass appeared in the culture tube. The cartilage lacuna was observed in the histological sections. Immunohistochemistry and in situ hybridization Positive. Conclusion Rabbit bone marrow mesenchymal stem cells can successfully produce cartilage tissue under the conditions of cartilage inducer and centrifuge tube culture.