2种不同成牙本质诱导液对大鼠脂肪干细胞增殖凋亡的影响

The effect of two different odontoblast inducer on the proliferation and apoptosis of rat adiposed-derived stem cells

目的探索2种不同的成牙本质诱导液——牙胚条件培养液(TGC-CM)和牙本质非胶原蛋白(DNCPM)对大鼠脂肪干细胞增殖和凋亡的影响。方法 (1)取出生4d的SD乳鼠4只,分别取腹股沟脂肪组织,用酶消化法进行脂肪干细胞分离和培养,免疫细胞化学法对细胞进行鉴定。(2)2种诱导液作用细胞后,倒置相差显微镜下观察细胞形态变化,MTT法检测细胞增殖情况,Annexin/PI双染后流式细胞仪检测细胞的凋亡情况。结果 (1)细胞培养至第2代时表型分子CD44和CD105表达呈阳性。(2)经TGC-CM及DNCPM培养3d后形态上发生极性改变。培养6d后,TGC-CM培养的细胞数量未见明显改变,而DNCPM培养的细胞密度明显降低。(3)MTT结果显示DNCPM组OD值低于对照组及TGC-CM组(P<0.05),流式细胞凋亡检测仪检测DNCPM组细胞凋亡率高于TGC-CM组和对照组(P<0.05)。结论作为大鼠脂肪干细胞向成牙本质样细胞方向分化的诱导液,TGC-CM可能更具优势。

Objective To investigate the effect of two different odontoblast inducer on the proliferation and apoptosis of rat adiposed-derived stem ceils. Methods Adiposed-derived stem cells were collected by enzyme digestion from inguinal fat pads of 4 days post natal mice. Immunocytochemistry was performed to identify the cells. MTT and flow cytometry were tested the prolifera- tion and apotosis of adipose＆derived stem cells by co-cultured with tooth germ cell conditioned medium（TGC-CM） or dentin non- collagenous protein medium （DNCPM）. Results Cells displayed a fibroblast-like appearance and positively expressed CD44 and CD105 when cufured to the secend yeneration. After 3 day the ceils polarity changed by co-cultured. Count of cells were no obvious change by TGC-CM co-cultured,while that ruduced significantly by DNCPM co-cultured. It confirmed that the proliferation rate of ADSCs in TGC-CM group and control group is higher than DNCPM group（P〈0.05） ,while the apotosis of that in DNCPM group is higher TGC-CM group（P〈0. 05）. Compared with control group, the results bad no significant difference（P〉0.05）. Conclusion TGC-CM may have more advantage as inducer in rat adiposed-derived stem cells differentiate into dentin like cells than DNCPM.