中国组织工程研究与临床康复2011，Vol.15Issue(14)：2592-2596,5.DOI:10.3969/j.issn.1673-8225.2011.14.026
大鼠阴茎海绵体肌源性干细胞的分离及鉴定
Isolation and determination of muscle-derived stem cells in the rat penile corpus cavernosum
许立军 1钟隆飞 1单玉喜 1薛波新 1陈岽1
作者信息
- 1. 苏州大学附属第二医院泌尿外科,江苏省苏州市,215004
- 折叠
摘要
Abstract
BACKGROUND: Previous studies have confirmed that stem cells with stem cell markers and muscle-derived stem cell markers were detected in rat penile corpus cavernosum.OBJECTIVE: To perform isolation and phenotype identification of muscle-derived stem cells in rat penile corpus cavernosum on the basis of previous study.METHODS: The penile corpus cavernosum from 2-month-old Sprague-Dawley rats was digested and dissociated by type Ⅰ collagenase and trypsin. Cells were primarily purified by the Preplate differential adhesion technique. PP1-PP6 adhered cells were obtained and the expressions of stem cell markers were detected by flow cytometry, immunofluorescence and western blotting.RESULTS AND CONCLUSION : The adherent capacity of PP1-PP6 cells was gradually weakened after 6-step differential adhesion. After two to three days, PP6 cells began to be adherent and became round or spindle-shaped. Flow cytometry detected stem cell antigen 1 (+) 5.7％, embryonic antigen (+) 2.6％, and desmin (+) 41.2％ in PP6 cells. Immunofluorescence exhibited that only a few cells expressed stem cell antigen 1 and embryonic antigen, showing diffused distribution. Stem cell antigen 1 was mainly expressed in cytoplasm, and embryonic antigen was mainly expressed in nuclei. Cell aggregation expressing desmin showed a large number and mainly expressed in cytoplasm. Very a few double-positive cells, such as stem cell antigen 1/embryonic antigen, stem cell antigen 1/desmin and embryonic antigen/desmin, were detected. No significant expression of stem cell antigen 1 and embryonic antigen protein was determined in PP1-PP5 cells. However, stem cell antigen 1/embryonic antigen protein was detected in PP6 cells. Desmin expression was gradually enhanced in PP1-PP6 cells. Results indicated that the cells from rat penile corpus cavernosum had the cell phenotype of stem cells and muscle-derived stem cells.关键词
肌源性干细胞/阴茎海绵体/干细胞抗原1/胚胎抗原/结蛋白/干细胞分类
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许立军,钟隆飞,单玉喜,薛波新,陈岽..大鼠阴茎海绵体肌源性干细胞的分离及鉴定[J].中国组织工程研究与临床康复,2011,15(14):2592-2596,5.
