| 毛囊bulge干细胞培养方法的比较研究 |
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| 引用本文： | 董刚,汪成林,彭栗,叶玲. 毛囊bulge干细胞培养方法的比较研究[J]. 华西口腔医学杂志, 2009, 27(6): 660-664. DOI: 10.3969/j.issn.1000-1182.2009.06.021 |
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| 作者姓名： | 董刚 汪成林 彭栗 叶玲 |
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| 作者单位： | 口腔疾病研究国家重点实验室,四川大学;口腔疾病研究国家重点实验室,四川大学;四川大学华西口腔医学院,牙体牙髓病教研室,四川,成都,610041 |
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| 基金项目： | 国家自然科学基金资助项目 |
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| 摘 要： | 目的通过对限定性角质形成细胞无血清培养基（DK-SFM）和3T3滋养层培养毛囊bulge干细胞的比较，寻求既能方便获得大量毛囊bulge干细胞，又能减少其分化的理想培养条件。方法采用DK-SFM和3T3滋养层分别培 养毛囊bulge干细胞，通过倒置显微镜观察毛囊bulge干细胞的形态，免疫荧光染色检测毛囊bulge干细胞的特异性标 记细胞角蛋白19（CK19）和分化相关抗体群34（CD34）的表达鉴定毛囊bulge干细胞。通过比较2种方法培养的毛囊bulge干细胞的克隆形成率和CD34阳性率的差异，分别比较毛囊bulge干细胞的增殖能力和干细胞的数量，综合评估 2种培养方法的优劣。结果倒置显微镜观察2种培养方法所得毛囊bulge干细胞均为铺路石状。免疫荧光染色检测 2种培养方法培养的毛囊bulge干细胞CK19和CD34均呈阳性。DK-SFM和3T3滋养层培养的毛囊bulge干细胞克隆形成 率分别为69.4%和62.2%。流式细胞仪检测DK-SFM培养的毛囊bulge干细胞中CD34阳性率为72.3%；3T3滋养层培养 的毛囊bulge干细胞中CD34阳性率为34.7％。结论DK-SFM和3T3滋养层培养作为毛囊bulge干细胞的2种培养方法， 均可以获得未分化的毛囊bulge干细胞。用3T3滋养层培养毛囊bulge干细胞的方法比较复杂，且所获的细胞混杂有 3T3细胞，但用此种方法可获得大量的毛囊bulge干细胞。用DK-SFM培养毛囊bulge干细胞方法简单，但细胞不易贴 壁，且数量较少，但能有效分离出较纯的毛囊bulge干细胞，保持较好的细胞活性。
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| 关 键 词： | 毛囊bulge干细胞 克隆形成率 细胞培养 |
| 收稿时间： | 2009-12-25 |
| 修稿时间： | 2009-12-25 |
Comparative study of cultivation of hair follicle bulge stem cell |
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| DONG Gang,WANG Cheng-lin,PENG Li,YE Ling. Comparative study of cultivation of hair follicle bulge stem cell[J]. West China journal of stomatology, 2009, 27(6): 660-664. DOI: 10.3969/j.issn.1000-1182.2009.06.021 |
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| Authors: | DONG Gang WANG Cheng-lin PENG Li YE Ling |
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| Affiliation: | 1. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China; 2. Dept. of Endodontics, West China College of Stomatology, Sichuan University, Chengdu 610041, China |
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| Abstract: | Objective The purpose was to compare two different ways of culturing hair follicle bulge stem cell: The defined keratinocyte-serum free medium(DK-SFM)method and the 3T3 feeder cell method.Methods The morphological features of cultured bulge stem cells were investigated by inverted phase control microscopy.Immunos-taining of stem cell marker cluster of differentiation 34(CD34)and epithelial cell marker cytokeratin 19(CK19)were performed to identify the bulge stem cell.The sternness of bulge stem cells was evaluated by colony forming efficiency(CFE)and proportion of CD34 positive cells by flow cytometry.Results Hair follicle bulge stem cells could be successfully cultivated in vitro using two methods.They were both positive for CK19 and CD34.The colony forming efficiency of hair follicle stem cell cultured in DK-SFM and the 3T3 feeder cell was 69.4% and 62.2%,respectively.There was no significant difference in colony forming efficiency between these two methods(P>0.05),while the CD34 positive cells proportion was higher in DK-SFM as 72.3% than the other as 34.7%(P<0.05).Conclusion Two methods are applicable to culture bulge stem cells in vitro.The 3T3 feeder cell method is complicated and can propagate a lot bulge stem cells from hair follicle,while the DK-SFM method is easier to get pure bulge stem cell. |
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| Keywords: | hair follicle bulge stem cell colony forming efficiency cell culture |
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