体外诱导猕猴骨髓基质干细胞向雪旺细胞分化  

Differentiation of rhesus bone marrow stromal cells into Schwann like cells in vitro

作　　者：胡军[1] 刘小林[1] 朱家恺[1] 邓宇斌[2] 向剑平[1] 易建华[1] 原清涛[2] 

机构地区：[1]中山大学附属第一医院骨科-显微外科医学部,广州市510080 [2]中山大学中山医学院病理生理教研室

出　　处：《中华创伤骨科杂志》2004年第5期538-541,共4页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金(30170962);广州市科技攻关重大项目(2002Z1-E0035)

摘　　要：目的探讨体外诱导猕猴骨髓基质干细胞向雪旺细胞分化的实验方法。方法采用BME、b-FGF、ATRA、BDNF、heregulin、Forskolin、PDGF依次联合诱导,倒置相差显微镜观察细胞形态学变化,免疫细胞化学鉴定S-100蛋白、GFAP和P75受体的表达率。结果诱导后的猕猴骨髓基质干细胞具有类似雪旺细胞的形态,免疫细胞化学结果显示S-100蛋白、GFAP和P75受体的表达率分别为(66.8±4.6)%、(57.3±5.4)%和(72.4±5.9)%。结论采用BME、ATRA、BDNF、heregulin、Forskolin、b-FGF、PDGF依次联合诱导,可使猕猴骨髓基质干细胞在体外向雪旺样细胞分化。Objective To determine the optimal protocol that can induce rhesus bone marrow stromal cells (BMSCs) to differentiate into Schwann like cells in vitro. Methods BMSCs were preinduced in a medium containing 1mm beta Mercaptoethanol (BME), 10ng/ml b FGF and 15%fetal bovine serum. After incubation for 24 hrs, cells were induced with all trans retinoic acid (ARTA) and BDNF for 4 to 6 hrs, followed by culture in the presence of heregulin, Forskolin, b FGF and PDGF for 5 days. The morphological changes positive of BMSCs were evaluated by inverted phase contrast microscope. The positive percentages of S100, GFAP and P75 expression were measured by immunocytochemistry with SABC staining. Results Rhesus BMSCs were positive for CD 29, CD 105, CD 166, and negative for CD 34, CD 71, CD 80 and CD 86. After the induction, BMSCs displayed the morphosis of Schwann cells, such as ellipsoidal cell bodies and fences like array with two or three slight cell processes. The percentages of positive S100, GFAP and P75 expression were 66.8%±4.6%, 57.3%±5.4%and 72.4%±5.9%respectively. Conclusion Rhesus MSCs can be induced to differentiate into Schwann like cells in vitro by BME, ATRA, BDNF, heregulin, Forskolin, b FGF and PDGF.

关 键 词：骨髓基质干细胞 细胞分化 雪旺细胞 猕猴 组织工程 周围神经缺损 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]