小鼠骨髓基质干细胞与脱细胞基质骨联合培养的实验研究

目的　研究小鼠股骨骨髓基质干细胞 (marrow stromal stem cells,MSCs)在同种异体脱细胞基质骨(acellular extra cellular matrix,AECM)上的生长情况。　方法　昆明种小鼠 8只共 16侧股骨 ,用 Triton X- 10 0处理后获 AECM。另将 1月龄昆明种小鼠 2 0只双侧股骨切断 ,用 4 ml PBS反复冲洗骨髓腔后 ,离心分瓶 ,取 MSCs进行原代培养 ,以 5× 10 6 /ml浓度将原代培养的小鼠 MSCs种植于 AECM中 ,进行体外联合培养 7d,通过相差显微镜、光镜、电镜等方法观察细胞在材料中的生长情况。　结果　在 AECM的孔隙中 ,有成团蓝染的 MSCs核团 ,周边部分大于中间部分 ,且靠近细胞培养液层面的 AECM中蓝染的核团数量明显比远离细胞培养液的层面多。 AECM具有良好的孔隙率 ,MSCs可在 AECM上良好生长。　结论　 AECM可作为构建组织工程骨的一种较好的支架材料。

STUDY ON TRANSPLANTATION OF MARROW STROMAL STEM CELLS INTO ACELLULAR EXTRA CELLULAR MATRIX IN RATS

OBJECTIVE: To study the integration of rat marrow stromal stem cells (MSCs) after transplantation into acellular extracellular matrix (AECM). METHODS: We got 16 femurs from 8 Kunming rats, the femurs were treated by Triton X-100 to get AECM, MSCs were collected from femoral marrow of 20 Kunming rats about a mouth old by PBS 4 ml, centrifuged and primary cultured in bottles, then the rat MSCs were transplanted into AECM at a concentration of 5 x 10(6)/ml and cultured for 7 days. The integration of the donor cells was observed using one phase contrast microscope, a light microscope and a scanning electron microscope (SEM). RESULTS: In AECM bone lacunas there were MSCs nuclei stained blue. The nuclei were unevenly distributed in AECM with more in the peripheral AECM than in the central AECM and with more in the layer near culture medium than in the layer far away from culture medium. AECM possessed a good spatial scaffold structure, the marrow stromal stem cells were well integrated into AECM. CONCLUSION: AECM can be used as a good scaffold material for tissue engineered bone construction.