不同浓度rhEPO对神经干细胞体外培养增殖的影响

The Influence of Different Density rhEPO to the Nerve Stem Cell in Vitro Raise Multiplication

目的观察不同浓度rhEPO对神经干细胞体外培养增殖的影响,探讨rhEPO联合神经干细胞移植治疗脊髓损伤修复的影响。方法用不同浓度(5、50、500 U/mL)rhEPO干预从新生SD大鼠(出生24 h以内)取材来的神经干细胞,分别检测每组神经干细胞克隆形成率;在不同时间分别用MTT法检测细胞增殖率;神经干细胞分化的免疫细胞化学染色及计数。结果添加rhEPO各实验组细胞增殖较快最终神经球的数量多于对照组,以50 U/mL rhEPO组作用显著;添加rhEPO各实验组的细胞生长曲线均高于对照组,尤其是50 U/mL rhEPO组显著高于对照组;NSE和GFAP免疫细胞化学染色和计数结果显示,50 U/mL rhEPO组中NSE免疫阳性细胞明显多于对照组(P<0.01)。结论rhEPO对神经干细胞体外培养增殖有促进作用,尤以适中浓度(50 U/mL)作用更加明显。

Objective To observe the effect which multiplies in vitro raise to the nerve stem cell of different density rhEPO,and to discuss the influence of rhEPO,associated with nerve stem cell transplant,in curing spinal cord damage repair.Methods The present research firstly intervened the nerve stem cell,obtained from the newborn SD big mouse（born within 24hs）,with different density rhEPO（5,50,500 U/mL）.Then,a distinctive examination of the rate of clone formation in each group＇s nerve stem cells was conducted.Afterwards,MTT method was used to examine the cell reproducibility in different times and nerve stem cell differentiation＇s immunocytochemistry dyeing and counting.Results The cell multiplication in experimental groups which had added the EPO was quicker and the quantity of final nerve balls was larger than that in the control group,which was remarkable in 50 U/mLEPO group. The cell growth curve in experimental groups which had added the rhEPO was higher than that in the control group,which was also remarkable in 50 U/mL EPO group.The NSE and GFAP immunocytochemistry dyeing and the counting results showed that the NSE immunity masculine cells in the 50 U/mL EPO group were obviously more than that in the control group（P0.01）.Conclusions rhEPO has the positive effect on the nerve stem cell in vitro raise multiplication,which is more obvious with the moderate density（50 U/ mL）.