" o8 t: m0 \" h/ t* @; F* S 【摘要】 为了探讨间充质干细胞(mesenchymal stem cell, MSC)诱导而来的内皮细胞的免疫调控能力，将人骨髓MSC在体外经VEGF诱导分化为内皮细胞，并用倒置相差显微镜和流式细胞术分别观察和检测细胞的形态和免疫表型，用淋巴细胞转化实验观察其免疫调控能力。结果表明： MSC在向内皮细胞分化的体系中生长迅速，诱导后的细胞经von Willebrand factor (vWF)免疫荧光染色呈阳性，能在Matrigel上形成毛细血管样网状结构。细胞表面标记无明显改变，即CD73、CD105、HLAABC仍阳性，CD34、 CD80、 CD86、 HLADR、 CD31仍为阴性。由MSC诱导分化而来的内皮细胞能抑制T淋巴细胞的增殖，其抑制作用随内皮细胞加入量的增多而增强。结论：由骨髓MSC来源的内皮细胞对T淋巴细胞具有调控作用。 % g) j5 H& F3 D
【关键词】间充质干细胞 内皮细胞 免疫调控 - g d* W3 S4 N Endothelial Cells Derived from Mesenchymal Stem Cells Harbor Immunoregulatory Effects2 g9 [, U3 ]; r7 t/ e3 ?3 P* k
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JIANG XiaoXia*, CHEN JinSong*1, SU YongFeng, LIAO Can1, LIU Bing, MAO Ning" Q+ |- Z4 W* F% ~5 }0 I# q2 a
- w7 A& I# [. e! F. u1 p6 a) F5 wInstitute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China; 1Guangzhou Cord Blood Bank, Guangzhou Maternal Neenatal Hospital, Guangzhou 510180, China) }2 E4 F. S# [4 w2 {
% F6 o7 e" m/ YAbstractThis study was purposed to investigate the immunoregulatory effect of endothelial cells derived from mesenchymal stem cells (MSC). The human MSC was induced to differentiate into endothelial cells for one week. The phenotypes were evaluated by flow cytometry,the cell morphologic feature was observed by invert phasecontrast microsoope and analysis of capillary formation was performed by using the in vitro angiogenesis kit. The immunoregulatory effect was detected bylymphocyte transformation test.The result indicatedthat during the differentiation cells grew fast and there was no significant change in the phenotypes, i.e.CD73，CD105，HLAABC were positiveand CD34, CD80, CD86, HLADR, CD31 were negative. Immunofluorescence analysis showed typical expression of the von Willebrand factor. Differentiated MSCs formed capillarylike structure. Endothelial cells derived from MSC also revealedimmunosuppressive effect on T cell proliferation in a dosedependent manner. It is concluded that endothelial cells derived from MSC also harbor immunoregulatory effecton T lymphocytes. 2 i0 b- P- a( J % U. k$ m3 L2 E* l: m( CKey wordsmesenchymal stem cell; endothelial cell; immunoregulatory effect ( H! N* p7 Z; k( F$ |0 l7 ]. h" o/ a- U0 y# {( Z
J Exp Hematol 2007; 15(1):175-178 ! Y' D) p" C" `: q& E+ U: Y+ t 7 U: t6 x2 {' `( z5 e! k ~1 F间充质干细胞(MSC)起源于中胚层，免疫原性低, 移植后无免疫排斥反应，对多种免疫细胞具有调控功能［1］，且在体外可大量增殖培养。若将MSC定向诱导，分化为血管内皮细胞，在体外构建成理想的血管移植物，在血管组织工程和再生医学领域必将有广阔的应用前景。5 d( R& ^; J0 K
$ [5 E: d5 A1 Z / F. h) y: M+ L) t: O- w3 k3 s 2Lyden D, Hattori K, Dias S, et al. Impaired recruitment of bonemarrowderived endothelial and hematopoietic precursor cells blocks tumor angiogenesis and growth. Nat Med, 2001; 7:1194-12014 K4 u% w, T$ D
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6 E' X8 }! g! y3 E: h4 _( p % ]3 Y9 N7 m3 ^ 3Jiang S, Walker L, Afentoulis M, et al. Transplanted human bone marrow contributes to vascular endothelium. Proc Natl Acad Sci USA, 2004; 101:16891-168961 `7 i5 \" P# B7 |
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4Shi Q, Rafii S, Wu MH, et al. Evidence for circulating bone marrowderived endothelial cells. Blood,1998; 92:362-367# a6 ~; A" r" |7 ?8 ?2 B7 d
: ~* }3 _/ U9 |4 q& e! K8 Q% V( N) O; F( R: s' b4 Z
( E, H. I; n/ z) C* I. \- I `- f 5Oswald J, Boxberger S, Jorgensen B, et al. Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells, 2004; 22:377-384 1 m* X2 D' T- k/ C7 {' G0 ]$ X+ M) f6 i n& s$ x
/ Q" v6 E6 D$ _7 O# @ & }: i. @5 x9 ^5 d& p" i% Q 6Muguruma Y, Yahata T, Miyatake H, et al. Reconstitution of the functional human hematopoietic microenvironment derived from human mesenchymal stem cells in the murine bone marrow compartment. Blood, 2006; 107:1878-1887 / ]5 L; {8 J2 c9 t# W, I' N+ K1 C. t; _6 d% r- a3 P9 L
. [7 ^# o+ q& B" _1 }& u0 r- I4 C. B2 c% @, n. U# x
7Reyes M, Dudek A, Jahagirdar B, et al. Origin of endothelial progenitors in human postnatal marrow. J Clin Invest, 2002; 109:337-346 ' m( G- K/ S2 q4 D , x9 L# v/ s3 e; {# b; h Q2 M+ f; X* F . _4 G( Z# v/ s u! m 8Asahara T, Murohara T, Sullivan A, et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science,1997; 275（5302）:964-967 - M# B$ ~1 R8 K& Z3 T7 C- N' I I* M8 k# w, g$ f
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4 k+ A2 ` N6 l) J+ V a# [ 9Gehling UM, Ergun S, Schumacher U, et al. In vitro differentiation of endothelial cells from AC133positive progenitor cells. Blood, 2000; 95:3106-3112 - _6 B. B8 c: r+ @) O3 ^, @8 w* Y1 w' F" C; s. T- R
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10LeBlanc K, Tammik C, Rosendahl K, et al. HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. Exp Hematol, 2003; 31:890-896［］μβαγ［AKU¨］ ［AKX］±SD作者: dypnr 时间: 2015-6-3 21:40