|本期目录/Table of Contents|

[1]侯红,吕安林△,达晶,等.人心脏干细胞向心肌细胞分化的研究[J].生物医学工程研究,2015,03:182-186.
　HOU Hong,LU Anlin,DA Jing,et al.The Study of Human Cardiac Stem Cells Differentiated into Cardiomyocytes in vitro[J].Journal of Biomedical Engineering Research,2015,03:182-186.
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人心脏干细胞向心肌细胞分化的研究(PDF)

《生物医学工程研究》[ISSN:1006-6977/CN:61-1281/TN]

期数:
2015年03期
页码:
182-186
栏目:
出版日期:
2015-09-25

文章信息/Info

Title:
The Study of Human Cardiac Stem Cells Differentiated into Cardiomyocytes in vitro
文章编号:
1672-6278 (2015)03-0182-05
作者:
侯红; 吕安林△; 达晶; 侯兆蕾
第四军医大学，西安市第二医院，西安 710003
Author(s):
HOU Hong; LU Anlin; DA Jing; HOU Zhaolei
Fourth Military Medical University,The Second Hospital of Xi ’an，Xi’an 710003, China
关键词:
人心脏干细胞; 分化; 心肌细胞; 5-氮杂胞苷; 血管紧张素 Ⅱ
Keywords:
Human cardial stem cells;  Differentiation;  Cardiomyocyte;  5-azacytidine;  Angiotensin Ⅱ
分类号:
R318
DOI:
-
文献标识码:
A
摘要:
研究了5-AZA、Ang II及其联合诱导人心脏干细胞向心肌细胞分化和提高分化率的方法。经患者或家属同意，从心外科手术中获取人心脏右心耳组织,Ⅱ型胶原酶消化、培养、传代，选用P2～P8细胞与心脏干细胞抗体CD117（c-kit）结合后，经流式细胞仪无菌分选纯化心脏干细胞，对分选纯化后的c-kit+CSCs进行培养（见前期试验）。 选取无菌分选纯化后c-kit+CSCs进行诱导、分化实验。实验随机分为四组：对照组（普通心脏干细胞培养液）、5-AZA诱导组、Ang II诱导组、5-AZA和Ang II联合诱导组。4周后用Western Blotting测定心肌细胞特异性蛋白cTn I、Cx43的表达；用流式细胞仪测定各组心肌细胞分化率。人心脏干细胞诱导4周后，Western blotting结果显示，四组均有cTn I、Cx43表达，对照组表达最弱，5-AZA和Ang II联合组表达最强。人心脏干细胞诱导4周后流式细胞仪测定各组心肌细胞分化率，统计结果显示：对照组(27.86±4.52) %、5-AZA组(52.71±7.40) %、Ang II组(40.48±7.02) %、5-AZA和Ang II联合组(64.74±6.11) %；四组间均有统计学差异（〖WTBX〗P〖WTBZ〗<0.05）。 5-AZA 、Ang II均能诱导人c-kit+CSCs分化为心肌细胞，5-AZA和Ang II联合诱导的心肌细胞分化率高于5-AZA、Ang II单独诱导方案。
Abstract:
To explore the differentiation of cardiac stem cells into cardiomyocytes and the methods of improving the differentiation rate of 5-AZA、II Ang and coinduced cardiac stem cells.Myocardial tissues obtained from the right atrial appendage of patients during surgery were digested with collagenaseⅡwith informed concent. The isolated cells were inoculated in culture flask and passaged, P2 to P8 cells were collected and incubated with monoclonal anti-human PE-CD117 specific antibody, then sterile sorting with flow cytometry. Then purified CSCs were collected for cultivation.After fusion around 90%, purified c-kit+CSCs were collected for differentiation and divided into four groups: （1）Control group(cells culture medium),（2）5-AZA group,（3）Ang-Ⅱgroup,（4）5-AZA combined Ang II group. The expression of protein of connexin 43 (Cx43) and cardiac Troponin I (cTn I) by Western Blot in 4 week and the cardiomyocyte differentiation rates with flow cytometry were determined.Identification of induced human CSCs cTn I, Cx43 protein expression: the expression of cTn I and Cx43 protein of each group by immunofluorescence staining was determined and the results showed that each group expressed protein of cTn I and Cx43, and weak expression had been found in the control group, strong expression in the 5-AZA combined Ang II group. Identification of induced human CSCs differentiation rates: the results detected by flow cytometry showed the cardiac differentiation rate of each group as follows: the control group was （27.86±4.52) %; the 5-AZA group was (52.71±7.40) %; the Ang II group was (40.48±7.02) %; the 5-AZA combined Ang II group was (64.74±6.11)%. Statistics of the differentiation rates between other groups had significant differences (〖WTBX〗P〖WTBZ〗<0.05).5-AZA, Ang II can induce cardiac stem cells to differentiate into cardiomyocyte, and the 5-AZA combined Ang II group is superior to single induced group.

参考文献/References

备注/Memo

备注/Memo:
(收稿日期：2015-06-08)通信作者Email:lüanlin@yahoo.com.cn
更新日期/Last Update: 2016-07-28