发育期牙根端复合体细胞条件培养液对脂肪干细胞增殖、分化的影响

发布时间：2018-08-26 07:56

【摘要】：目的:初步探讨发育期根端复合体(developing apical complex,DAC)条件培养基(DAC conditioned medium,DACCM)对脂肪干细胞(adipose tissue-derived stem cells,ADSCs)增殖和分化的影响。方法:取出生20 d SD大鼠分离其下颌磨牙DAC,并采用组织块联合酶消化法培养DAC细胞;经HE、免疫组化染色鉴定DAC后,收集原代培养的DAC细胞上清,分别用过滤和不过滤的方式制备不同条件培养基并用其对ADSCs进行培养;分别于培养3、5、7 d各时间点,MTT法检测ADSCs增殖能力;用碱性磷酸酶(ALP)试剂盒及反转录酶-聚合酶链反应(RT-PCR)检测ADSCs的ALP蛋白活性及其基因的表达水平。结果:免疫组化染色显示,培养的DAC细胞CK-14、vimentin均为阳性表达;DACCM-未滤培养组ADSCs的增殖活性、ALP蛋白活性以及ALP基因表达水平均高于单纯α-MEM培养组和DACCM-过滤培养组,差异均有统计学意义(P0.05)。结论:不经过滤的DAC细胞上清制备的条件培养基对ADSCs增殖、分化的促进作用更明显。
[Abstract]:Aim: to investigate the effect of (DAC conditioned medium,DACCM on the proliferation and differentiation of adipose stem cells (adipose tissue-derived stem cells,ADSCs). Methods: the DAC, of mandibular molars was isolated from 20 day old SD rats and DAC cells were cultured by tissue mass and enzyme digestion. The supernatants of DAC cells were collected after DAC was identified by HE, immunohistochemical staining. ADSCs was cultured in different condition culture medium by filtration and unfiltration, and the proliferation ability of ADSCs was detected by MTT assay at each time point of 57-day culture. Alkaline phosphatase (ALP) kit and reverse transcriptase polymerase chain reaction (RT-PCR) were used to detect ALP protein activity and gene expression of ADSCs. Results: the results of immunohistochemical staining showed that the positive expression of CK-14,vimentin in DAC cells was positive, and the activity of ADSCs and the expression of ALP gene were higher in cultured DAC cells than those in 伪 -MEM culture group and DACCM- filtration culture group. The difference was statistically significant (P0.05). Conclusion: the conditioned medium prepared by the supernatant of unfiltered DAC cells can promote the proliferation and differentiation of ADSCs.
【作者单位】：安徽医科大学口腔医学院;安徽医科大学附属口腔医院牙周科安徽省口腔疾病研究中心实验室;安徽医科大学人兽共患病安徽省重点实验室;
【基金】：安徽省自然科学基金资助项目(1308085MH130)
【分类号】：R781.4

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