作者：何伟， 沈铁城，狄荣科，杨勇，张志坚作者单位：(江苏大学附属医院骨科， 江苏 镇江 212001) - L: z, b! r& d 9 b) m9 U+ c: m+ C& i
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【摘要】 目的： 探讨褪黑激素（melatonin，MT）体外诱导大鼠骨髓间充质干细胞(marrow mesenchymal stem cells，MSCs)不同时段向神经细胞分化的情况。方法： 采用细胞培养技术分离培养和纯化大鼠MSCs，以MT为诱导剂诱导大鼠MSCs向神经细胞分化。分别在诱导后3，7，10，14 d以免疫荧光法测定神经干细胞特异性标志(nestin) 、神经元细胞特异性标志(neurofilament, NF)和星形胶质细胞特异性标志(GFAP)的表达。结果： MT诱导分化3 d，部分细胞胞体收缩成三角形,有些成为简单的双极或多极细胞。诱导3 d细胞开始表达nestin, 7～14 d表达 nestin和NF,细胞不表达GFAP。结论： MT可诱导大鼠MSCs分化为神经细胞。 ; k/ U- j6 k5 ~9 c# I& ~ 【关键词】骨髓间充质干细胞； 褪黑激素； 神经细胞( [. q) H/ ^8 Y/ c& T$ q
In vitro melatonin induction of differentiation of marrow" w! z" s0 A) N Q
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HE Wei, SHEN Tiecheng, DI Rongke, YANG Yong, ZHANG Zhijian+ v) L" [# K# G% ?
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(Department of Orthopedics, Affiliated Hospital, Jiangsu University, Zhenjiang Jiangsu 212001,China) , w+ g1 q" I3 Y, |" G' j) Y, M : [% x4 c. }4 i: T6 L7 ^ ［Abstract］Objective: To explore MT introduce marrow mesenchymal stem cells into neurons on different time. Methods： By using cell culture method, rat MSCs were isolated and purified, The rat MSCs were induced with MT.The expression of nestin,NF and GFAP in the differentiated cells was detected by immunofluorescence after introduced 3,7,10,14 days.Results： Three days after induction with MT some neurons were found to contract, refract more light and protrude and bulge.Some neurons contracted to atriangular shape, and some others became simple bipolar or multipolar cells. After being induced for 3 days, the cells expressed nestin.After 7～14 days, the cells expressed nestin and NF, but didn′t express GFAP.Conclusion： MT can introduce MSCs into neurons.* B4 U) V: ?, G! x1 ^* k
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【参考文献】 ! e2 j z$ c6 h* ^7 d+ z$ P ［1］Long X, Olszewski M, Huang W, et al. Neural cell differentiation in vitro from adult human bone marrow mesenchymal stem cells ［J］.Stem Cells Dev,2005,14（1）:65-69. ! t3 H* [) m" |4 }6 ] F# ? 9 i8 U) b% _; A$ B5 O | B. T- B$ M2 O4 e! J0 r2 F * h3 a( H, j) K/ F& A3 R5 |) ~ ［2］Mimura T, Dezawa M, Kanno H, et al. Peripheral nerve regeneration by transplantation of bone marrow stromal cellderived Schwann cells in adult rats ［J］. J Neurosurg, 2004, 101(5):806-812.0 r% L$ X3 J4 v7 W6 {
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- A6 B; {4 A3 P; t: B2 J$ p ［3］Dezawa M. Insights into autotransplantation: the unexpected discovery of specific induction systems in bone marrow stromal cells ［J］. Cell Mol Life Sci, 2006, 63(23):2764-2772. # z! P9 t l. c$ L' b+ ]) ` ( H# H1 j3 T( T- ^ 2 T" {9 ]9 C" ?8 b/ C- W ) ]+ D1 t0 n/ ?# R' Z ［4］Woodbury D,Schwarz EJ,Prockop DJ,et al.Adult rat and human bone marrow stromal cells differentiate into neurons［J］.J Neurosci Res, 2000, 61(4): 364-370. A5 s* F. b- i3 Y; y/ N" B/ |& E