兔骨髓间质干细胞的体外分离、培养及活性鉴定

Isolating and culturing rabbit bone marrow stromal cells in vitro and identifying the activity

目的：建立一种体外分离培养自体兔骨髓间质干细胞（ Ｍ ａｒｒｏｗ Ｓｔｒｏｍ ａｌ Ｃｅｌｌｓ， Ｍ Ｓ Ｃｓ）的方法。方法：抽取兔骨髓液３～５ ｍ ｌ，经密度梯度离心得骨髓单个核细胞，以１６×１０４／ｃｍ ２ 细胞浓度培养，１２～１４ ｄ 后得到贴壁生长的单层细胞，随后进入传代培养，每传一代约 ５～７ ｄ。结果：原代培养的、以及传代后培养的细胞均为贴壁生长的、均匀一致的纺锤形细胞。两种细胞经体外特殊环境诱导后均可转化为骨髓网状间质细胞和成骨细胞。结论：该细胞经体外长期培养后仍具有多潜能转化活性，确系骨髓间质干细胞。为该细胞的广泛应用奠定基础。

Objective:To establish a method of isolating and chulturing rabbit bone marrow stromal cells (MSCs)in vitro.Methods:A 3~5 ml bone marrow was extracted from the iliac of Japanese white rabbits.The nucleated cells fraction of the marrow was enriched based on density centrifuge and culatured by 1 6×10 4 cells/cm 2 The adherent cells approached confluence 12~14 days after plating and then every passage was cultured for 5~7 days.Results:The adherent cells during the first time and passaged several times in culture became more uniformly spindle shaped in appearance.They both might be induced to differentiate into bone marrow mesenchymal cells and osteoblast under special environment.Conclusion:The method for isolating and culturing rabbit MSCs was feasible,and these cells still possess the ability to differentiate into multiple cells after passage several times.