目的分离培养大鼠毛囊Bulge细胞并观察其生物学特性。方法运用显微分离法及酶消化法分离大鼠毛囊Bulge细胞并进行培养,观察其增殖能力及细胞形态等的改变;采用免疫组化方法,检测毛囊Bulge细胞K19、岛整合素的表达情况。结果本方法培养的大鼠毛囊Bulge细胞纯度高、活力好,具有高增殖、低分化特性,同时表达K19、岛整合素。结论成功地分离培养了毛囊Bulge细胞,并在体外长期保持其低分化特性,为进一步深入研究毛囊发育分化提供了实验基础。
Objective To isolate and culture the hair follicle Bulge cells of rats and study their morphological, immunological characteristics. Methods Hair follicle Bulge obtained by micromanipulation and enzyme digestion was cultured in intro. The morphological features and numbers of Bulge cells were identified and counted with light microscopy. Immunocytochemical staining was used to detect expressing of K19 and β1 integrin in cultured cells. Results Bulge cells after enzyme digestion pasted rapidly. Cells were small and round, paving stone-shape, and had a large ratio of nuclear to cytoplasm. The characteristics suggested a primitive morphologic feature. Cells growth well and could maintain low differentiation and higher reproductive activity, they co-expressing K19 and β1 integrin, immunofluorescence staining showed the cells expressed a6-integrin strongly, weakly or no expressing CD71. Conclusion This culture system can amplify a lot of pure hair follicle Bulge cells in short time. We successfully isolate and culture the hair follicle Bulge cells of rats in intro and can keeping its property for a long time.