我们的网站为什么显示成这样？

可能因为您的浏览器不支持样式，您可以更新您的浏览器到最新版本，以获取对此功能的支持，访问下面的网站，获取关于浏览器的信息：

|本期目录/Table of Contents|

生长分化因子-11促进小鼠诱导性多能干细胞向心肌细胞定向分化的研究

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2016年第3期
页码:
279-284
栏目:
基础研究
出版日期:
2016-01-05

文章信息/Info

Title:
Growth differentiation factor 11 promotes cardiac differentiation of mouse-induced pluripotent stem cells
作者:
郭 涛1; 刘 通1; 陈江炜1; 江振华1; 李秀娟1; 樊苗苗1; 曹 丰2
(1.第四军医大学西京医院心内科，陕西 西安 710032；
2.中国人民解放军总医院心内科,北京 100853)
Author(s):
GUO Tao1;  LIU Tong1;  CHEN Jiang-wei1;  JINAG Zhen-hua1;  LI Xiu-juan1;  FAN Miao-miao1;  CAO Feng2
(1.Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi, China；
2.Department of Cardiology, PLA General Hospital, Beijing 100853, China)
关键词:
生长分化因子-11; 诱导多能干细胞; 细胞分化; 小鼠
Keywords:
growth differentiation factor-11;  mouse-induced pluripotent stem cells;  cardiac differentiation;  mouse
分类号:
R392.12
DOI:
-
文献标识码:
A
摘要:
目的 明确生长分化因子（GDF）-11对小鼠诱导多能干细胞（miPSCs）向心肌细胞定向分化的促进作用，为心肌再生的细胞生物学治疗提供种子细胞和实验依据。方法 常规培养小鼠miPSCs,分为对照组和GDF-11组。GDF-11组在普通分化培养基中加用10 ng/ml的GDF-11。悬滴法诱导培养形成拟胚体（EBs），每日观察搏动拟胚体数目。采用实时荧光定量PCR检测多能干细胞标志物Oct-4、心脏中胚层标志物Flk-1、心脏祖细胞标志物Nkx2.5、和心肌特异性标志物cTnT的表达变化。用免疫荧光染色观察心肌结构蛋白cTnI的表达水平。结果 GDF-11组的Oct-4表达水平在诱导分化的第3、7天分别为同期对照组的（0.55±0.31）倍（P<0.01）和（0.41±0.57）倍（P<0.05）；诱导分化的第10天，GDF-11组的Flk-1和Nkx2.5表达相分别为对照组的（2.09±0.8）倍（P<0.05）和（2.47±0.22）倍（P<0.01）；cTnT的表达在分化后第10天和第14天分别为对照组的（1.81±0.19）倍（P<0.01）和（1.61±0.20）倍（P<0.01）；两组均出现了心肌样搏动细胞团，GDF-11组搏动EBs百分比要显著高于对照组（P<0.01）。免疫荧光染色显示，GDF-11组的cTnI阳性率要显著高于对照组（P<0.01）。结论 GDF-1能够显著促进miPSCs的心肌定向分化。
Abstract:
AIM To study the effect of growth differentiation factor 11 (GDF-11) on the cardiac differentiation of mouse-induced pluripotent stem cells (miPSCs) and to provide promising cells and an experimental basis for the cellular biological therapy of cardiomyocyte regeneration. METHODS MiPSCs were cultured using conventional methods and were divided into GDF-11 group and control group. The differentiation medium of the GDF-11 group contained 10 ng/ml GDF-11. Embryoid bodies (EBs) were formed using the hanging drop method and the number of beating EBs was calculated daily. Expression of pluripotent stem cell marker Oct-4, cardiac mesoderm marker Flk-1, cardiac progenitor marker Nkx2.5 and cardiac specific marker cTnT were detected using qRT-PCR. The expression of cTnI was observed using immunoflourensence staining. RESULTS Expression levels of Oct-4 in GDF-11 group at day 3 and day 7 were (0.55±0.31)-fold (P<0.01) and (0.41±0.57)-fold (P<0.05) of those in the control group. The expressions of flk-1 and Nkx2.5 in GDF-11 group at day 10 were higher (2.09±0.8)-fold (P<0.05) and (2.47±0.22)-fold (P<0.01) than the control group. In addition, GDF-11 group at days 10 and 14 had a (1.81±0.19)-fold change (P<0.01) and a (1.61±0.20)-fold change (P<0.01) of cTnT expression compared with those in the control group. Beating areas were found in both groups, but GDF-11 group had a higher percentage of beating EBs. Immunoflourensence staining showed more cTnI positive cells in the GDF-11 group (P<0.01). CONCLUSION GDF-11 significantly enhances cardiac differentiation of miPSCs.

参考文献/References

[1]Go AS,Mozaffarian D,Roger VL,et al.Heart disease and stroke statistics--2013 update:a report from the American Heart Association[J].Circulation,2013,127(1):e6-e245.
[2]Takahashi K,Yamanaka S.Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J].Cell,2006,126(4):663-676.
[3]Mauritz C,Schwanke K,Martin U,et al.Generation of functional murine cardiac myocytes from induced pluripotent stem cells[J].Circulation,2008,118(5):507-517.
[4]Yan B,Abdelli LS,Singla DK,et al.Transplanted induced pluripotent stem cells improve cardiac function and induce neovascularization in the infarcted hearts of db/db mice[J].Mol Pharm,2011,8(5):1602-1010.
[5]Ott MG,Marme F,Seimetz D,et al.Humoral response to catumaxomab correlates with clinical outcome: results of the pivotal phase II/III study in patients with malignant ascites[J].Int J Cancer,2012,130(9):2195-2203.
[6]Vaidya A,Kale VP.TGF-beta signaling and its role in the regulation of hematopoietic stem cells[J].Syst Synth Biol,2015,9(1-2):1-10.
[7] Watabe T,Miyazono K.Roles of TGF-beta family signaling in stem cell renewal and differentiation[J].Cell Res,2009,19(1):103-115.
[8]Jha R,Xu RH,Xu C.Efficient differentiation of cardiomyocytes from human pluripotent stem cells with growth factors[J].Methods Mol Biol,2015,1299:115-131.
[9]Cagavi E,Bartulos O,Qyang Y,et al.Functional cardiomyocytes derived from Isl1 cardiac progenitors via Bmp4 stimulation[J].PLoS One,2014,9(12):e110752.
[10]Takei S,Ichikawa H,Sasaki K,et al.Bone morphogenetic protein-4 promotes induction of cardiomyocytes from human embryonic stem cells in serum-based embryoid body development[J].Am J Physiol Heart Circ Physiol,2009,296(6):H1793-H1803.
[11]Loffredo FS,Steinhauser ML,Lee RT,et al.Growth differentiation factor 11 is a circulating factor that reverses age-related cardiac hypertrophy[J]. Cell,2013,153(4):828-839.
[12]Yu J,Vodyanik MA,Thomson JA,et al.Induced pluripotent stem cell lines derived from human somatic cells[J].Science,2007,318(5858):1917-1920.
[13]Dimos JT,Rodolfa KT,Eggan K,et al.Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons[J].Science,2008,321(5893):1218-1221.
[14]Park IH,Arora N,Daley GQ,et al.Disease-specific induced pluripotent stem cells[J].Cell,2008,134(5):877-886.
[15]Narazaki G,Uosaki H,Yamashita JK,et al.Directed and systematic differentiation of cardiovascular cells from mouse induced pluripotent stem cells[J].Circulation,2008,118(5):498-506.
[16]Bock C,Kiskinis E,Meissner A,et al.Reference Maps of human ES and iPS cell variation enable high-throughput characterization of pluripotent cell lines[J].Cell,2011.144(3):439-452.
[17]Kim K,Doi A,Daley GQ,et al.Epigenetic memory in induced pluripotent stem cells[J].Nature,2010,467(7313):285-290.
[18]Kattman SJ,Witty AD,Keller G,et al.Stage-specific optimization of activin/nodal and BMP signaling promotes cardiac differentiation of mouse and human pluripotent stem cell lines[J].Cell Stem Cell,2011,8(2):228-40.
[19]Laflamme MA,Chen KY,Murry CE,et al.Cardiomyocytes derived from human embryonic stem cells in pro-survival factors enhance function of infarcted rat hearts[J].Nat Biotechnol,2007,25(9):1015-1024.
[20]Sinha M,Jang YC,Wagers AJ,et al.Restoring systemic GDF11 levels reverses age-related dysfunction in mouse skeletal muscle[J].Science,2014,344(6184):649-652.
[21]Zadegan G,Ghaedi K,Kalantar SM,et al.Cardiac differentiation of mouse embryonic stem cells is influenced by a PPAR gamma/PGC-1alpha-FNDC5 pathway during the stage of cardiac precursor cell formation[J].Eur J Cell Biol,2015,94(6):257-266.
[22]Jha R,Xu RH,Xu C.Effect of miR-20b on Apoptosis, Differentiation,the BMP Signaling Pathway and Mitochondrial Function in the P19 Cell Model of Cardiac Differentiation In Vitro[J].PLoS One,2015,10(4):e0123519.
[23]Kerscher P,Bussie BS,Lipke EA,et al.Characterization of mitochondrial populations during stem cell differentiation[J].Methods Mol Biol,2015,1264:453-463.
[24]Hochedlinger K,Yamada Y,Jaenisch R,et al.Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues[J].Cell,2005,121(3):465-477.
[25]Prall OW,Menon MK,Harvey RP,et al.An Nkx2-5/Bmp2/Smad1 negative feedback loop controls heart progenitor specification and proliferation[J].Cell,2007,128(5):947-959.

备注/Memo

备注/Memo:
收稿日期：2015-06-02.
基金项目：军队十二五重点项目资助（BWS12J037）；国家杰出青年基金项目资助（81325009）
通讯作者：曹丰，教授，主要从事干细胞分子影像学研究 Email：wind8828@gmail.com
作者简介：郭涛，硕士生 Email：guotao19871228@163.com
更新日期/Last Update: 2016-01-07