中文摘要：

目的 探讨剥夺钙和血清对培养的骨髓基质细胞（BMSCs）的影响.方法 将传至第2代的BMSCs用DMEM培养液和无钙DMEM培养液按不同比例配制成细胞悬液,细胞浓度为2＆#215;104 mL-1,再根据是否加10%FBS分为6组：无10%FBS正常钙组（1组）、10%FBS正常钙组（2组,正常对照组）、10%FBS1/3正常钙组（3组）、10%FBS1/6正常钙组（4组）、10%FBS无钙组（5组）和无10%FBS无钙组（6组）.培养后4 h,1-7 d用CCK-8试剂盒检测BMSCs细胞增殖数.无10%FBS无钙组、无10%FBS正常钙组、10%FBS无钙组和10%FBS正常钙组在培养后0、4、12、24和48 h,0、1、3、5和7 d 时在倒置相差显微镜下观察贴壁生长BMSCs细胞形态的变化.结果 1组、6组培养后4 h,1-7 d 时细胞增殖数均明显低于2组（均P＜0.01）,3组培养后4 h,3-5 d时细胞增殖数明显低于2组（P＜0.05或P＜0.01）;4组培养后4 h,1、3和5 d时细胞增殖数均显著低于2组（均P＜0.01）,培养后7 d时细胞增殖数显著高于2组（P＜0.01）;5组培养后4 h,1-3、5 d时细胞增殖数均显著低于2组（均P＜0.01）.10%FBS无钙组、10%FBS正常钙组细胞形态正常,继续贴壁生长、增殖,细胞密度增加;无10%FBS无钙组、无10%FBS正常钙组细胞培养后4 h时开始贴壁细胞变小,培养后1、3 d时大量细胞漂浮、死亡,仅有极少数细胞保持贴壁.结论剥夺钙和血清均对细胞生存不利,低钙浓度对于BMSCs有短期抑制生存作用,但BMSCs能适应低钙环境,恢复活力和增殖.BMSCs适应低钙的意义与机制有待进一步探讨.

英文摘要：

Objective To investigate the effects of calcium and serum deprivation on bone mar row stromal cells （BMSCs） in vitro. Methods The second passage BMSCs were suspended in DMEM with or without calcium at a concentration of 2 X 10^4 cells/mL. The suspended cells were divided into six groups：serum deprivation and normal calcium level group（group 1）, 10% serum and normal calcium level group（group 2, normal control group）, 10 % serum and 1/3 levels of nor- mal calcium group（group 3）, 10% serum and 1/6 levels of normal calcium group（group 4）, 10% serum and calcium-free group（group 5）,and serum deprivation and calcium-free group（group 6）. The number of living cells was counted using Cell Counting Kit-8 at 4 hours and 1-7 days after treatment. In addition,cells in group 1,2,5 and 6 were grown in fresh medium after culture for 6 days. The changes in morphology of adherent BMSCs were observed under an inverted phase con-trast microscope at 0,4,12,24 and 48 hours and 0,1,3,5 and 7 days after medium replacement. Results Compared with group 2, the number of cells decreased in group 1 at 4 hours and 1 7 days,group 3 at 4 hours and 3-5 days,group 4 at 4 hours and 1,3 and 5 days and group 5 at 4 hours and 1,2,3 and 5 days after treatment, but increased in group 4 at 7 days after treatment （P〈0.05 or P〈0.01）. Cells in group 2 and 5 had normal morphology and continue to proliferate in an adherent fashion with an increase in cell density. In group 1 and 6, cells shrank after culture for 4 hours and a large number of floating dead cells were found after culture for 1,3 days with very few cells remaining attached to the plate. Conclusion Calcium and serum deprivation was detrimental to the survival of BMSCs. Low calcium culture exerts a short-term inhibition on the survival of BMSCs,but BMSCs can adapt to low calcium environment,restore viability and main- tain proliferation. However,the significance and mechanism need to be further explored.