Generation and Characterization of Functional Cardiomyocytes from Rhesus Monkey Embryonic Stem Cells
Generation and Characterization of Functional Cardiomyocytes from Rhesus Monkey Embryonic Stem Cells
复制标题
恒河猴胚胎干细胞功能性心肌细胞的生成和表征
DOI：
10.1634/stemcells.2005-0380
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发表时间：
2006
期刊：
影响因子：
5.2
通讯作者：
U. Martin
中科院分区：
文献类型：
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作者：
K. Schwanke;S. Wunderlich;M. Reppel;M. Winkler;Matthias Matzkies;S. Groos;J. Itskovitz‐Eldor;A. Simon;J. Hescheler;A. Haverich;U. Martin
来自小鼠和人类的胚胎干细胞（hESCs）已被证明能够有效地向心肌细胞（CMs）分化。由于小鼠ESCs和hESCs不允许建立临床前同种异体移植模型，我们研究的目的是从恒河猴ESCs （rESCs）中产生功能性CMs。尽管外胚层和神经元/胶质细胞的形成似乎是rESC系R366.4的默认途径，但我们能够改变这一承诺，直接产生内胚层/中胚层细胞，并进一步向CMs分化。rESCs的分化导致平均18%的rESCs自发收缩胚状体（EBs）。半定量逆转录-多聚酶链反应分析显示，在rESC来源的CMs中，内胚层、中胚层、心脏中胚层和CMs中典型的标记基因表达，包括brachyury、goosecoid、Tbx‐5、Tbx‐20、Mesp1、Nkx2.5、GATA‐4、FOG‐2、Mlc2a、MLC2v、ANF和α‐MHC。免疫组织学和超微结构研究显示CM典型蛋白的表达，包括肌动蛋白、肌钙蛋白T、titin、连接蛋白43和交叉横纹肌原纤维。通过多电极阵列进行的电生理学研究揭示了生成的CMs的功能、电偶联和β -肾上腺素能信号传导的证据。这是第一个证明从rESCs衍生出功能性CMs的研究。与hESCs相比，rESCs允许建立临床前同种异体移植模型。此外，rESC衍生的CMs为心脏安全药理学的高通量测定提供了一个细胞来源。
Embryonic stem cells (ESCs) from mice and humans (hESCs) have been shown to be able to efficiently differentiate toward cardiomyocytes (CMs). Because murine ESCs and hESCs do not allow for establishment of pre‐clinical allogeneic transplantation models, the aim of our study was to generate functional CMs from rhesus monkey ESCs (rESCs). Although formation of ectodermal and neuronal/glial cells appears to be the default pathway of the rESC line R366.4, we were able to change this commitment and to direct generation of endodermal/mesodermal cells and further differentiation toward CMs. Differentiation of rESCs resulted in an average of 18% of spontaneously contracting embryoid bodies (EBs) from rESCs. Semiquantitative reverse transcription‐polymerase chain reaction analyses demonstrated expression of marker genes typical for endoderm, mesoderm, cardiac mesoderm, and CMs, including brachyury, goosecoid, Tbx‐5, Tbx‐20, Mesp1, Nkx2.5, GATA‐4, FOG‐2, Mlc2a, MLC2v, ANF, and α‐MHC in rESC‐derived CMs. Immunohistological and ultrastructural studies showed expression of CM‐typical proteins, including sarcomeric actinin, troponin T, titin, connexin 43, and cross‐striated muscle fibrils. Electrophysiological studies by means of multielectrode arrays revealed evidence of functionality, electrical coupling, and β‐adrenergic signaling of the generated CMs. This is the first study demonstrating generation of functional CMs derived from rESCs. In contrast to hESCs, rESCs allow for establishment of pre‐clinical allogeneic transplantation models. Moreover, rESC‐derived CMs represent a cell source for the development of high‐throughput assays for cardiac safety pharmacology.
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作者：
K-Y Francis Pau;D. Wolf;K-Y Francis Email;P. Edu ; Don;Wolf
通讯作者：
K-Y Francis Pau;D. Wolf;K-Y Francis Email;P. Edu ; Don;Wolf
DOI：
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作者：
Biochemistr;Biochemistr And Biochemistry;M. Halbach;U. Egert;J. Hescheler;K. Banach
通讯作者：
Biochemistr;Biochemistr And Biochemistry;M. Halbach;U. Egert;J. Hescheler;K. Banach
DOI：
10.1073/pnas.92.17.7844
发表时间：
1995-08-15
影响因子：
11.1
作者：
THOMSON, JA;KALISHMAN, J;HEARN, JP
通讯作者：
HEARN, JP
DOI：
10.1073/pnas.95.15.8801
发表时间：
1998-07-21
影响因子：
11.1
作者：
Kajstura, J;Leri, A;Anversa, P
通讯作者：
Anversa, P
DOI：
10.1152/ajpheart.1997.272.1.h220
发表时间：
1997-01-01
影响因子：
4.8
作者：
Soonpaa, MH;Field, LJ
通讯作者：
Field, LJ