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人胚胎干细胞系 KIND-2 向具有多能性的心血管祖细胞的分化。

Differentiation of human ES cell line KIND-2 to yield tripotent cardiovascular progenitors.

机构信息

Stem Cell Biology Department, National Institute for Research in Reproductive Health, Parel, Mumbai, 400 012, India.

出版信息

In Vitro Cell Dev Biol Anim. 2013 Jan;49(1):82-93. doi: 10.1007/s11626-012-9558-0. Epub 2013 Jan 4.


DOI:10.1007/s11626-012-9558-0
PMID:23288411
Abstract

Human embryonic stem cells (hESCs) have the ability to differentiate into all the three lineages and are an ideal starting material to obtain cells of desired lineage for regenerative medicine. Continued efforts are needed to evolve more robust protocols to obtain cells of desired lineages and in larger numbers. Also, it has now been realized that rather than transplanting fully committed cells differentiated in vitro, it may be ideal to transplant committed progenitors which retain the intrinsic ability to proliferate and also differentiate better into multiple lineages based on the in vivo cues. For cardiac regeneration, the desired progenitor is a multipotent cardiovascular progenitor which has the ability to regenerate cardiomyocytes, endothelial cells, and also smooth muscle cells. The present study was undertaken to carefully compare three widely used protocols to differentiate hESCs into cardiac progenitors, viz., spontaneous differentiation, differentiation by END-2-conditioned medium, and directed differentiation using growth factors followed by quantitative PCR to study the relative expression of early cardiovascular markers. hESC differentiation mimicked the early embryonic development, and the transition into mesoendoderm, mesoderm, early cardiac progenitors, and cardiac cells associated with spontaneous beating was clearly evident in all the three groups. However, compared to spontaneous and END-2-associated differentiation, directed differentiation led to several-fold higher expression of cardiac transcripts (>75-fold Nkx2.5 and >150-fold Tbx5) in response to the stage-specific addition of well-established cardiogenic inducers and inhibitors of specific signaling pathways. We propose to use tripotent cardiovascular progenitors derived by directed differentiation for further preclinical studies.

摘要

人胚胎干细胞(hESCs)具有分化为所有三个谱系的能力,是获得所需谱系细胞的理想起始材料,用于再生医学。需要继续努力制定更强大的方案,以获得所需谱系的更多数量的细胞。此外,现在已经意识到,与其移植体外分化的完全分化细胞,不如移植具有内在增殖能力的定向祖细胞,并根据体内线索更好地分化为多个谱系,可能更为理想。对于心脏再生,所需的祖细胞是多能心血管祖细胞,具有再生心肌细胞、内皮细胞和平滑肌细胞的能力。本研究旨在仔细比较三种广泛使用的将 hESC 分化为心脏祖细胞的方案,即自发分化、END-2 条件培养基分化和使用生长因子进行定向分化,然后进行定量 PCR 研究早期心血管标志物的相对表达。hESC 分化模拟了早期胚胎发育,并在所有三组中都明显过渡到中胚层、中胚层、早期心脏祖细胞和与自发跳动相关的心脏细胞。然而,与自发分化和 END-2 相关分化相比,定向分化导致心脏转录物的表达增加了数倍(>75 倍 Nkx2.5 和>150 倍 Tbx5),这是由于对特定阶段添加了成熟的心脏诱导剂和特定信号通路抑制剂。我们建议使用定向分化获得的三潜能心血管祖细胞进行进一步的临床前研究。

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引用本文的文献

[1]
Transcriptional activator DOT1L putatively regulates human embryonic stem cell differentiation into the cardiac lineage.

Stem Cell Res Ther. 2018-4-10

[2]
Evaluating KIND1 human embryonic stem cell-derived pancreatic progenitors to ameliorate streptozotocin-induced diabetes in mice.

Indian J Med Res. 2017-8

[3]
Shifting gears from embryonic to very small embryonic-like stem cells for regenerative medicine.

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[4]
Different Tissue-Derived Stem Cells: A Comparison of Neural Differentiation Capability.

PLoS One. 2015-10-30

[5]
Stem cells, progenitors & regenerative medicine: A retrospection.

Indian J Med Res. 2015-2

[6]
Pluripotent stem cells for cardiac regeneration: overview of recent advances & emerging trends.

Indian J Med Res. 2013-2

本文引用的文献

[1]
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Stem Cells Dev. 2012-6-13

[2]
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Nat Biotechnol. 2011-10-23

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Stem Cells Int. 2011-9-6

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Cell Stem Cell. 2011-8-5

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Induction of MesP1 by Brachyury(T) generates the common multipotent cardiovascular stem cell.

Cardiovasc Res. 2011-6-1

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Evaluating differentiation propensity of in-house derived human embryonic stem cell lines KIND-1 and KIND-2.

In Vitro Cell Dev Biol Anim. 2011-5-26

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Induced pluripotent stem cell (iPSC)-derived Flk-1 progenitor cells engraft, differentiate, and improve heart function in a mouse model of acute myocardial infarction.

Eur Heart J. 2011-5-19

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PLoS One. 2011-4-8

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