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JoVE Journal
Developmental Biology
从胚胎干细胞衍生心脏祖细胞
从胚胎干细胞衍生心脏祖细胞
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
Derivation of Cardiac Progenitor Cells from Embryonic Stem Cells

从胚胎干细胞衍生心脏祖细胞

Full Text
8,651 Views
08:00 min
January 12, 2015

DOI: 10.3791/52047-v

Ieng Lam Lei1, Lei Bu2, Zhong Wang1

1Cardiac Surgery,University of Michigan, 2Leon H Charney Division of Cardiology,New York University School of Medicine

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol demonstrates the derivation of cardiac progenitor cells from both mouse and human embryonic stem cells. The process involves enriching these cells using flow cytometry with fluorescent reporters.

Key Study Components

Area of Science

  • Cardiac biology
  • Stem cell research
  • Cell differentiation

Background

  • Embryonic stem cells (ESCs) are a valuable resource for generating specific cell types.
  • Cardiac progenitor cells are crucial for heart development and repair.
  • Flow cytometry allows for the precise isolation of desired cell populations.
  • Understanding the differentiation process can aid in regenerative medicine.

Purpose of Study

  • To generate cardiac progenitor cells from embryonic stem cells.
  • To establish a reliable protocol for cell enrichment and differentiation.
  • To facilitate further research into cardiac cell therapies.

Methods Used

  • Culturing high-quality embryonic stem cells.
  • Transferring cells to gelatin-coated dishes for embryo body formation.
  • Adding growth factors to induce further differentiation.
  • Dissociating embryo bodies and treating with additional growth factors.

Main Results

  • Successful generation of cardiac progenitor cells from ESCs.
  • Establishment of a flow cytometry-based enrichment method.
  • Potential for further differentiation into cardiomyocytes and smooth muscle cells.
  • Demonstration of a reproducible protocol for cardiac cell generation.

Conclusions

  • This protocol provides a framework for deriving cardiac progenitor cells.
  • It highlights the importance of growth factors in cell differentiation.
  • The method can be adapted for various research applications in cardiac biology.

Frequently Asked Questions

What are cardiac progenitor cells?
Cardiac progenitor cells are precursors to heart cells that can differentiate into cardiomyocytes and smooth muscle cells.
Why is flow cytometry used in this protocol?
Flow cytometry is used to enrich cardiac progenitor cells by isolating specific populations based on fluorescent markers.
Can this protocol be used for human embryonic stem cells?
Yes, the protocol is designed to work with both mouse and human embryonic stem cells.
What role do growth factors play in this process?
Growth factors are essential for promoting the differentiation of embryonic stem cells into cardiac progenitor cells.
What are the potential applications of this research?
This research can contribute to regenerative medicine and therapies for heart diseases.

在该协议中,将说明来自小鼠和人胚胎干细胞的心脏祖细胞的衍生。该方案中的一个主要策略是使用工程化到胚胎干细胞系中的荧光报告基因,通过流式细胞术富集心脏祖细胞。

该程序的总体目标是从胚胎干细胞生成人或小鼠心脏祖细胞。这是通过首先在高质量的胚胎干细胞或 e ESC 或小鼠胚胎饲养层细胞或 myth 中培养来实现的。在第二步中,将细胞转移到明胶涂层的培养皿中,以促进第一个胚胎体的形成。

接下来,将细胞关联并将生长因子添加到培养物中以诱导第二种形式的胚胎体。在最后一步中,胚胎体再次解离,并用额外的生长因子处理所得细胞。最终,心脏后代细胞可以进一步培养用于心肌细胞和平滑肌细胞分化。

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