大鼠组织提取液对体外培养骨髓基质细胞的影响

The effect of tissue fluid extracted on bone marrow stromal cells cultured in vitro

目的研究大鼠脑组织成份和其它组织提取液对体外培养骨髓基质细胞(bonemarrowstromalcells,BMSC)的影响。方法提取大鼠全脑、灰质、白质、垂体、肌肉、肝脏等组织液,进行原代骨髓基质细胞培养,观察并测定其ALP染色阳性率。以空白培养液作对照,观察大鼠各组织提取液对骨髓基质细胞增殖,分化,矿化的作用。结果(1)各组织提取液对原代骨髓基质细胞培养后,其ALP染色阳性率以灰质和全脑组最高,其次是白质和肌肉组。(2)除肝脏组外其余各组织提取液对BMSC均有不同程度的增殖刺激作用,以灰质最强,其它依次为肌肉、白质、全脑与血清组,肝脏组则显示对BMSC增殖有抑制作用。(3)肝脏组对BMSC具有分化抑制作用,其它各组均显示分化刺激作用,增加程度依次为灰质、全脑、白质、肌肉与血清。(4)灰质组的矿化结节形成率最高,其它依次为全脑、白质、肌肉与血清。结论大鼠脑组织中的灰质提取液能显著刺激原代骨髓基质细胞增殖,且向成骨细胞诱导分化,并且在二代以后的BMSC培养中,表现出最强的刺激增殖、向成骨细胞分化和体外矿化的能力。

Objective Study the effect of tissue fluid extracted from the rat brain components and other tissue on bone marrow stromal cells (BMSC) cultured in vitro. Methods Tissue fluid was extracted from rat brain, white matter, gray matter, hypophysis, liver and muscle, respectively. Primary rat BMSC were cultured in vitro with various types of tissue extract. ALP staining was done and the effect on BMSC proliferation, differentiation, and mineralization was observed. BMSC cultured with culture media only served as control. Results (1) ALP positive staining rate after primary BMSC culture was found to be highest with gray matter extract and with brain extract. BMSC cultures with white matter extract and muscle extract yielded next highest ALP positive staining rate. (2) Extract of all the tissues except liver could stimulate BMSC proliferation, the gray matter extract being the highest followed by muscle extract, white matter extract, brain extract and serum. Liver extract inhibited BMSC proliferation. (3) Liver extract inhibited BMSC differentiation, while all other tissue extract stimulated BMSC differentiation, their activity in descending order being gray matter extract, brain extract, white matter extract, muscle extract and serum. (4) The highest mineralization nod formation rate was found in culture with gray matter extract, followed by culture with brain extract, white matter extract, muscle extract and serum. Conclusion Gray matter tissue extract can significantly stimulate proliferation of the primary BMSC and induce it to become osteoblast. It also has capability of stimulating BMSC proliferation, differentiation and mineralization.