兔骨折血肿细胞的体外培养及诱导分化

Expansion of Rabbit Fracture Hematoma Cells in vitro

目的 建立一种体外培养骨折血肿细胞方法,探讨血肿细胞的成骨潜力。 方法 体外分离、纯化兔骨折血肿细胞,置于3种不同培养液中,观察比较细胞生长情况;用条件培养液诱导血肿细胞定向分化,检测细胞的碱性磷酸酶和骨钙素的表达。 结果 (1)经3种不同培养液培养的兔骨折血肿细胞在P1和P2 时生长和增殖情况无明显差别,P4以后生长速度有明显差异(P <0 .0 1) ;(2 )血肿细胞在αMEM培养液中增殖活跃,诱导分化后细胞碱性磷酸酶为阳性,对照组为阴性,少量为弱阳性;经条件培养液培养的血肿细胞骨钙素含量高于未加诱导剂培养的细胞(P <0 .0 1)。 结论 兔骨折血肿细胞在体外合适的条件下,生长稳定,有较强的增殖能力,具有肯定的成骨能力。

Objective To establish a method for culturing the fracture hematoma cells in vitro and investigate the osteogenic capability of rabbit fracture hematoma cells. Methods Rabbit fracture hematoma cells were isolated, purified and cultured in three different culture mediums and the proliferation and growth characteristics were observed and compared; the osteogenic potential of cultured hematoma cells in a conditional medium were examined by histochemistry stains technique. Results The growth and proliferation of rabbit fracture hematoma cells in three different culture mediums were not significantly different in the first and second passage culture, while they showed decreased after the fourth passage culture(P<0.01). The hematoma cells had strong capability of proliferation in α-MEM medium, and the derived cells showed positive staining of ALP. The quantity of osteocalcin(OCN) was higher in α-MEM medium than in the other medium cultured. Conclusion Under appropriate in vitro conditions, rabbit fracture hematoma cells keep active proliferative capacity in primary and passage cultures.