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背景:骨髓中存在非黏附干细胞,该类细胞形态及表型特征与黏附骨髓间充质干细胞相似。目的:观察胎源性非黏附骨髓基质细胞的基本特性。方法:采用反复贴壁分离法培养胎源性骨髓非黏附基质细胞,每24h取悬浮细胞分瓶培养,共4次;采用MTT法测定第1,3,5代细胞生长曲线,流式细胞仪行细胞周期测定和表面标志分析;并取第1代细胞行造血集落培养,第3代细胞成骨和成脂诱导培养。结果与结论:反复贴壁分离培养4次后仍有细胞贴壁并有集落形成,剩余细胞经培养7d后即可得到原代骨髓非黏附基质细胞。骨髓非黏附基质细胞形态不均匀,高表达整合素及内皮细胞表面标记,中等量表达CD106,同时也表达造血细胞标记CD34;体外可分化为成骨细胞及造血细胞,但成脂分化能力较弱。
BACKGROUND: Non-adherent stem cells exist in bone marrow. The morphological and phenotypic characteristics of these cells are similar to those of adherent BMSCs. Objective: To observe the basic characteristics of fetal non-adherent bone marrow stromal cells. Methods: The fetal bone marrow non-adherent stromal cells were cultured by repeated adherent separation method. The suspension cells were cultured for 4 times every 24h, and the growth curve of cells on day 1, 3 and 5 were measured by MTT assay. The flow cytometry Line cell cycle determination and surface marker analysis; and take the first generation of cells hematopoietic line colony culture, third generation osteoblast and adipogenic induction. RESULTS AND CONCLUSION: The cells still adhered and colonized after repeated adherent culture for 4 times. The remaining cells were cultured for 7 days to obtain the primary bone marrow non-adherent stromal cells. Bone marrow non-adherent stromal cells inhomogeneous morphology, high expression of integrin and endothelial cell surface markers, moderate expression of CD106, also expressed hematopoietic cell marker CD34; in vitro can be differentiated into osteoblasts and hematopoietic cells, but adipogenic differentiation is weak .