BMSCs 向心肌分化的基因表達譜分析_《中國修復重建外科雜志》

作者：

楊琳 ¹ , 沈悌 ² , 陳連鳳 ² , 曹欣欣 ² , 賴晉智 ³

1 華中科技大學同濟醫學院附屬同濟醫院腫瘤科（武漢，430030）;;
中國醫學科學院北京協和醫學院北京協和醫院 2 血液內科，3 心內科;

關鍵詞：

hBMSCs 基因表達譜誘導心肌樣細胞

視頻：

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目的分析hBMSCs 經5- 氮雜胞苷（5-azacytidine，5-aza）誘導向心肌樣細胞分化過程中基因表達譜
的改變。方法取胸外科非血液病患者手術中廢棄的肋骨骨髓分離培養hBMSCs，5-aza 誘導第2 代hBMSCs。取誘導
前及誘導后的細胞，免疫細胞化學法檢測α-actin、肌鈣蛋白T（cardiac troponin T，cTnT）和連接蛋白43（connexin 43）表
達，流式細胞儀檢測cTnT 陽性細胞百分率；利用人表達譜基因芯片技術篩選分化過程中的差異表達基因，對部分基因進
行功能分類和分層聚類分析。結果 hBMSCs 經5-aza 誘導后，部分呈肌細胞樣形態。免疫細胞化學染色檢測示誘導
前細胞α-actin、cTnT 染色呈弱陽性，connexin 43 染色呈陰性；誘導后3 周細胞α-actin、cTnT、connexin 43 染色均呈陽性。
流式細胞儀檢測示誘導前cTnT 陽性細胞百分率為7.43% ± 0.02%，誘導后3 周為49.64% ± 0.05%。分化過程中共檢測到
1 814 個顯著差異表達基因，對其中647 個基因分層聚類，聚為5 類，生物功能包括信號傳導、細胞代謝、增殖分化、發育以
及形態發生等。結論 hBMSCs 經5-aza 誘導向心肌樣細胞分化過程受信號傳導通路、轉錄基因、生長因子等多種因素
在不同時間點上的共同調控。

引用本文： 楊琳 ,沈悌,陳連鳳,曹欣欣,賴晉智. BMSCs 向心肌分化的基因表達譜分析. 中國修復重建外科雜志, 2012, 26(5): 607-611. doi: 復制

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3.	Britten MB, Abolmaali ND, Assmus B, et al. Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction (TOPCARE-AMI): mechanistic insights from serial contrast-enhanced magnetic resonance imaging. Circulation, 2003, 108(18): 2212-2218.
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8.	Cohen ED, Tian Y, Morrisey EE. Wnt signaling: an essential regulator of cardiovascular differentiation, morphogenesis and progenitor self-renewal. Development, 2008, 135(5): 789-798.
9.	Flaherty MP, Abdel-Latif A, Li Q, et al. Noncanonical Wnt11 signaling is sufficient to induce cardiomyogenic differentiation in unfractionated bone marrow mononuclear cells. Circulation, 2008, 117(17): 2241-2252.
10.	Qyang Y, Martin-Puig S, Chiravuri M, et al. The renewal and differentiation of isl1+ cardiovascular progenitors are controlled by a Wnt/beta-catenin pathway. Cell Stem Cell, 2007, 1(2): 165-179.
11.	Zhu W, Shiojima I, Ito Y, et al. IGFBP-4 is an inhibitor of canonical Wnt signalling required for cardiogenesis. Nature, 2008, 454(7202): 345-349.
12.	Schneider VA, Mercola M. Wnt antagonism initiates cardiogenesis in Xenopus laevis. Genes Dev, 2001, 15(3): 304-315.
13.	Tzahor E, Lassar AB. Wnt signals from the neural tube block ectopic cardiogenesis. Genes Dev, 2001, 15(3): 255-260.
14.	Li Y, Hiroi Y, Ngoy S, et al. Notch1 in bone marrow-derived cells mediates cardiac repair after myocardial infarction. Circulation, 2011, 123(8): 866-876.
15.	Crispino JD, Lodish MB, Thurberg BL, et al. Proper coronary vascular development and heart morphogenesis depend on interaction of GATA-4 with FOG cofactors. Genes Dev, 2001, 15(7): 839-844.
16.	Zhao R, Watt AJ, Battle MA, et al. Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. Dev Biol, 2008, 317(2): 614-619.
17.	Sachinidis A, Kolossov E, Fleischmann BK, et al. Generation of cardiomyocytes from embryonic stem cells experimental studies. Herz, 2002, 27(7): 589-597.
18.	Yang HS, Bhang SH, Kim IK, et al. In situ cardiomyogenic differentiation of implanted bone marrow mononuclear cells by local delivery of transforming growth factor-β1. Cell Transplant, 2011. [Epub ahead of print].
19.	Wan CR, Chung S, Kamm RD. Differentiation of embryonic stem cells into cardiomyocytes in a compliant microfluidic system. Ann Biomed Eng, 2011, 39(6): 1840-1847.
20.	Afouda BA, Hoppler S. Different requirements for GATA factors in cardiogenesis are mediated by non-canonical Wnt signaling. Dev Dyn, 2011, 240(3): 649-662.

1. Passier R, van Laake LW, Mummery CL. Stem-cell-based therapy and lessons from the heart. Nature, 2008, 453(7193): 322-329.
2. Wollert KC, Meyer GP, Lotz J, et al. Intracoronary autologous bone-marrow cell transfer after myocardial infarction: the BOOST randomised controlled clinical trial. Lancet, 2004, 364(9429): 141-148.
3. Britten MB, Abolmaali ND, Assmus B, et al. Infarct remodeling after intracoronary progenitor cell treatment in patients with acute myocardial infarction (TOPCARE-AMI): mechanistic insights from serial contrast-enhanced magnetic resonance imaging. Circulation, 2003, 108(18): 2212-2218.
4. Dimmeler S, Zeiher AM, Schneider MD. Unchain my heart: the scientific foundations of cardiac repair. J Clin Invest, 2005, 115(3): 572-583.
5. Ling SK, Wang R, Dai ZQ, et al. Pretreatment of rat bone marrow mesenchymal stem cells with a combination of hypergravity and 5-azacytidine enhances therapeutic efficacy for myocardial infarction. Biotechnol Prog, 2011, 27(2): 473-482.
6. Xu W, Zhang X, Qian H, et al. Mesenchymal stem cells from adult human bone marrow differentiates into a cardiomyocyte phenotype in vitro. Exp Biol Medi (Maywood), 2004, 229(7): 623-631.
7. Makino S, Fukuda K, Miyoshi S, et al. Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest, 1999, 103(5): 697-705.
8. Cohen ED, Tian Y, Morrisey EE. Wnt signaling: an essential regulator of cardiovascular differentiation, morphogenesis and progenitor self-renewal. Development, 2008, 135(5): 789-798.
9. Flaherty MP, Abdel-Latif A, Li Q, et al. Noncanonical Wnt11 signaling is sufficient to induce cardiomyogenic differentiation in unfractionated bone marrow mononuclear cells. Circulation, 2008, 117(17): 2241-2252.
10. Qyang Y, Martin-Puig S, Chiravuri M, et al. The renewal and differentiation of isl1+ cardiovascular progenitors are controlled by a Wnt/beta-catenin pathway. Cell Stem Cell, 2007, 1(2): 165-179.
11. Zhu W, Shiojima I, Ito Y, et al. IGFBP-4 is an inhibitor of canonical Wnt signalling required for cardiogenesis. Nature, 2008, 454(7202): 345-349.
12. Schneider VA, Mercola M. Wnt antagonism initiates cardiogenesis in Xenopus laevis. Genes Dev, 2001, 15(3): 304-315.
13. Tzahor E, Lassar AB. Wnt signals from the neural tube block ectopic cardiogenesis. Genes Dev, 2001, 15(3): 255-260.
14. Li Y, Hiroi Y, Ngoy S, et al. Notch1 in bone marrow-derived cells mediates cardiac repair after myocardial infarction. Circulation, 2011, 123(8): 866-876.
15. Crispino JD, Lodish MB, Thurberg BL, et al. Proper coronary vascular development and heart morphogenesis depend on interaction of GATA-4 with FOG cofactors. Genes Dev, 2001, 15(7): 839-844.
16. Zhao R, Watt AJ, Battle MA, et al. Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. Dev Biol, 2008, 317(2): 614-619.
17. Sachinidis A, Kolossov E, Fleischmann BK, et al. Generation of cardiomyocytes from embryonic stem cells experimental studies. Herz, 2002, 27(7): 589-597.
18. Yang HS, Bhang SH, Kim IK, et al. In situ cardiomyogenic differentiation of implanted bone marrow mononuclear cells by local delivery of transforming growth factor-β1. Cell Transplant, 2011. [Epub ahead of print].
19. Wan CR, Chung S, Kamm RD. Differentiation of embryonic stem cells into cardiomyocytes in a compliant microfluidic system. Ann Biomed Eng, 2011, 39(6): 1840-1847.
20. Afouda BA, Hoppler S. Different requirements for GATA factors in cardiogenesis are mediated by non-canonical Wnt signaling. Dev Dyn, 2011, 240(3): 649-662.

《中國修復重建外科雜志》

BMSCs 向心肌分化的基因表達譜分析

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《中國修復重建外科雜志》

BMSCs 向心肌分化的基因表達譜分析

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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