人羊膜细胞与创伤性脑组织提取液共培养的实验研究

目的 探究将人羊膜细胞(HACs)置于创伤性脑组织提取液的条件培养基中,模拟HACs在创伤性脑损伤微环境下的分化和增殖情况,为HACs脑内移植提供基础研究.方法 取无并发症剖官产羊膜,用0.25%胰酶反复消化3次提取HACs,接种后48h去除未贴壁的HACs.采用改进的Feeney法制作大鼠创伤性脑损伤(TBI)模型,TBI 24 h后制备创伤性脑组织提取液.将HACs分别接种于RPMI 1640培养基(1640)、RPMI 1640培养基与正常脑组织提取液(1640 NB)、RPMI 1640培养基与创伤件脑组织提取液(1640 TBI)3组.共培养7 d后HE染色显微镜下检测各组细胞形态.培养前后分别行Nestin和MAP-2单抗免疫组化检测.将HACs以104/ml密度分别接种上述3种培养基,培养后1、4和7 d经酶联免疫检测仪行四氮唑盐(MTT)比色法检测各组HACs活性.结果 1640组HACs呈圆形或椭圆形,核大居中;1640 TBI组部分HACs形态出现锥形、三角形等神经元样改变.HACs培养前可见Nestin阳性细胞,培养后1640 TBI组可见MAP-2阳性表达.各组HACs增殖性均不佳.结论 HACs与创伤性脑组织提取液共培养后可转化为神经元样细胞.HACs增殖性不佳.

An experimental study on human amniotic cells co-culture by traumatic brain tissue extracts

Objective To explore the differentiation and proliferation of human amniotic cells(HACs) in the conditioned culture medium with traumatic brain tissue extracts,which mimics the microenvironment of injured brain.Methods A human amniotic sheet was obtained from an uncomplicated elective Cesarean section,and then the separated amnion was treated with 0.25% trypsin for three times to collect HACs within an hour.A rat model of traumatic brain injury(TBI) was established by improved Feeney's technique,and the tra...