培养骨髓基质细胞的羟脯氨酸-2-表异构酶活性

CYTOCHEMICAL OBSERVATION OF THE EPIMERASE ACTIVITIES OF BONE MARROW STROMAL CELLS IN CULTURE

本实验利用液体静置法体外培养小鼠骨髓基质细胞.对生长细胞进行倒置显微镜下的原位连续观察及数种细胞化学观察.结果发现,骨髓游离细胞在两周内逐日减少,而贴壁细胞逐日增殖和分化.在成纤维细胞,网状细胞和巨噬细胞中均出现很强的羟脯氨酸-2-表异构酶(Hydro-xyproline-2-epimerase,EC5.1.1.a简称HP-Ⅱ-E)活性.证明杂环亚氨基酸-羟脯氨酸参与这些细胞代谢的过程非常活跃.

Fibroblasts, reticular cells and macrophages are the important components of bone marrow hemopoietic in liquid to study whether HP-Ⅱ-E(Hydroxyproli- ne-2-epimerase, EC5, 1, 1,a) is involved in the synthesis of functional proteins of stromal cells. The femoral bone of 6 to 8 week-old 615 mice were made in suspension for cell culture in vitro. DMEM medium supplied with 25% calf serum was used After one to fifteen day's attaching incubation in stationary liquid the culture cells were stained with Giemsa, nonspecific esterase and HP-Ⅱ-E at different intervals. From the third day of the incubation, reticular cells began to develop form- ing stallate cells of the bone marrow. During an earlier time, enzymatic reaction was only observed in the nuclear membrance, but after three day's culture all the enzymatic activity in the cells increased dramatically. The enzymatic reac- tive granules were abundant in the outer cytoplasma of the stallate cells and fibroblasts and the cytoplasma of the macrophages. It was observed that hyd- roxyproline was changed by epimerase,