大鼠牙髓干细胞的培养和鉴定

目的：分离培养大鼠牙髓干细胞并对其进行生物学鉴定。方法：采用单细胞克隆的方法进行大鼠牙髓干细胞的分离培养,并将达到一定数量级的牙髓干细胞与HA—TCP支架材料进行复合,移植入裸鼠背部皮下,8周后取材进行组织学鉴定。取鉴定成功的细胞株复苏进行细胞生物学特性研究,同时进行成牙本质细胞诱导并进行DsP、DMP1免疫组织化学染色一结果：单细胞来源的细胞克隆(RDPSC282)移植物组织切片可见牙本质牙髓复合体样结构。复苏细胞形态为梭形,细胞密集时形态多样,为卵圆形或多角形,细胞群体倍增时间为58．3h,克隆形成率为1．33％,免疫组化染色DSP、DMPl均为15月性,经矿化液诱导后部分细胞胞浆DsP及DMP1出现阳性染色。结论：首次成功分离培养出一株大鼠牙髓干细胞。

Culture and identification of rat dental pulp stem cell

AIM:To isolate,culture and identify a rat dental pulp stem cell line.METHODS:Single cell clones of enzymatically isolated adult rat dental pulp cells were culture-expanded and seeded into HA/TCP bioceramics to generate a cell-scaffold construct,which was then implanted subcutaneously into the dorsal side of nude mice.Samples were extracted 8 weeks post-implantation for histological examination. The cell clones that demonstrated dentin-pulp complex structure formation during in vivo study were recovered from liquid nitrogen and taken for odontogenic induction,which was identified by immunohistochemical staining of DSP and DMP1.RESULTS:Implants seeded with cell clone named RDPSC2B2 demonstrated dentin-pulp complex-like structure formation.The recovered cells from this clone were generally fusiform,and vary in morphology from ellipse to polygonal when grown to confluence.The cells were stained positive by DSP and DMP1 after induced by mineralizing culture medium. The doubling time of the cell is 58.3 hours and clone formation unit is 1.33%.CONCLUSION:We isolated a rat dental pulp stem cell line.