人胚干细胞源性 CD133+内皮祖细胞移植治疗性血管生成修复心肌。

Therapeutic angiogenesis by transplantation of human embryonic stem cell-derived CD133+ endothelial progenitor cells for cardiac repair.

机构信息

Falk Cardiovascular Research Centre, Stanford School of Medicine, USA.

出版信息

Regen Med. 2010 Mar;5(2):231-44. doi: 10.2217/rme.09.83.

DOI:10.2217/rme.09.83

PMID:20210583

Abstract

OBJECTIVE

This study aim to enhance endothelial differentiation of human embryonic stem cells (hESCs) by transduction of an adenovirus (Ad) vector expressing hVEGF(165) gene (Ad-hVEGF(165) ). Purified hESC-derived CD133(+) endothelial progenitors were transplanted into a rat myocardial infarct model to assess their ability to contribute to heart regeneration.

METHODS

Optimal transduction efficiency with high cell viability was achieved by exposing differentiating hESCs to viral particles at a ratio of 1:500 for 4 h on three consecutive days.

RESULTS

Reverse transcription-PCR analysis showed positive upregulation of VEGF, Ang-1, Flt-1, Tie-2, CD34, CD31, CD133 and Flk-1 gene expression in Ad-hVEGF(165) -transduced cells. Additionally, flow cytometric analysis of CD133, a cell surface marker, revealed an approximately fivefold increase of CD133 marker expression in Ad-hVEGF(165)-transduced cells compared with the nontransduced control. Within a rat myocardial infarct model, transplanted CD133(+) endothelial progenitor cells survived and participated, both actively and passively, in the regeneration of the infarcted myocardium, as seen by an approximately threefold increase in mature blood vessel density (13.62 +/- 1.56 vs 5.11 +/- 1.23; p < 0.01), as well as significantly reduced infarct size (28% +/- 8.2% vs 76% +/- 5.6%; p < 0.01) in the transplanted group compared with the culture medium-injected control. There was significant improvement in heart function 6 weeks post-transplantation, as confirmed by regional blood-flow analysis (1.72 +/- 0.612 ml/min/g vs 0.8 +/- 0.256 ml/min/g; p < 0.05), as well as echocardiography assessment of left ventricular ejection fraction (60.855% +/- 7.7% vs 38.22 +/- 8.6%; p < 0.05) and fractional shortening (38.63% +/- 9.3% vs 25.2% +/- 7.11%; p < 0.05).

CONCLUSION

hESC-derived CD133(+) endothelial progenitor cells can be utilized to regenerate the infarcted heart.

摘要

目的

本研究旨在通过转导表达人 VEGF(165) 基因的腺病毒（Ad）载体增强人胚胎干细胞（hESC）的内皮分化。纯化的 hESC 衍生的 CD133(+)内皮祖细胞被移植到大鼠心肌梗死模型中，以评估其促进心脏再生的能力。

方法

通过将分化的 hESC 暴露于病毒颗粒，比例为 1:500，连续 3 天，每天 4 小时，实现了具有高细胞活力的最佳转导效率。

结果

逆转录-PCR 分析显示，Ad-hVEGF(165)转导的细胞中 VEGF、Ang-1、Flt-1、Tie-2、CD34、CD31、CD133 和 Flk-1 基因表达呈阳性上调。此外，通过流式细胞术分析 CD133 这一细胞表面标志物，与未转导的对照组相比，Ad-hVEGF(165)转导的细胞中 CD133 标志物的表达增加了约 5 倍。在大鼠心肌梗死模型中，移植的 CD133(+)内皮祖细胞存活下来，并积极和被动地参与梗死心肌的再生，成熟血管密度增加了约 3 倍（13.62±1.56 对 5.11±1.23；p<0.01），移植组的梗死面积明显减小（28%±8.2%对 76%±5.6%；p<0.01）。与对照组相比，移植后 6 周时心脏功能明显改善，通过区域性血流分析得到证实（1.72±0.612 ml/min/g 对 0.8±0.256 ml/min/g；p<0.05），以及超声心动图评估左心室射血分数（60.855%±7.7%对 38.22%±8.6%；p<0.05）和缩短分数（38.63%±9.3%对 25.2%±7.11%；p<0.05）。