Dong Gang, Wang Cheng-Lin, Peng Li, Ye Ling
State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2009 Dec;27(6):660-4.
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.
The morphological features of cultured bulge stem cells were investigated by inverted phase control microscopy. Immunostaining 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 stemness of bulge stem cells was evaluated by colony forming efficiency (CFE) and proportion of CD34 positive cells by flow cytometry.
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).
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.
比较两种不同的毛囊隆突干细胞培养方法：限定角质形成细胞无血清培养基（DK-SFM）法和3T3饲养层细胞法。
通过倒置相差显微镜观察培养的隆突干细胞的形态特征。进行干细胞标志物分化抗原34（CD34）和上皮细胞标志物细胞角蛋白19（CK19）的免疫染色以鉴定隆突干细胞。通过集落形成效率（CFE）和流式细胞术检测CD34阳性细胞比例来评估隆突干细胞的干性。
两种方法均可成功在体外培养毛囊隆突干细胞。它们的CK19和CD34均呈阳性。在DK-SFM和3T3饲养层细胞中培养的毛囊干细胞的集落形成效率分别为69.4%和62.2%。这两种方法之间的集落形成效率无显著差异（P>0.05），而DK-SFM中CD34阳性细胞比例更高，为72.3%，另一种为34.7%（P<0.05）。
两种方法均适用于体外培养隆突干细胞。3T3饲养层细胞法操作复杂，但能从毛囊中扩增大量隆突干细胞，而DK-SFM法更容易获得纯化的隆突干细胞。
