Improve Research Reproducibility A Bio-protocol resource
Peer-reviewed

Mouse Embryonic Stem Cell Differentiation to Hematopoietic Precursors

小鼠胚胎干细胞(ES)分化造血前体

HI Hogune Im

发布: 2012年04月05日第2卷第7期 DOI: 10.21769/BioProtoc.144 浏览次数: 15575

download pdf PDF PDF
ask Q&A
引用
收藏
Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of The Journal of Biological Chemistry, featuring study using the protocol.

May 2003

Presubmission Inquiry

实验方案合集

专注于特定技术或应用的、详细的、经过同行评审的实验方案合集

Cancer Research
Immunology
Developmental Biology
Microbiology
Neuroscience
Cell Imaging - A Special Collection for Cell Bio 2023
查看更多 more

相关实验方案

小鼠原始内胚层干细胞的产生

小鼠原始内胚层干细胞的产生

Yasuhide Ohinata [...] Haruhiko Koseki

2023年11月20日 1759 阅读

用于单细胞RNA测序的人骨髓非造血细胞的分离

用于单细胞RNA测序的人骨髓非造血细胞的分离

Hongzhe Li [...] Stefan Scheding

2024年06月20日 1665 阅读

通过CRISPR介导的Tp53基因敲除高效永生化小鼠胚胎成纤维细胞

通过CRISPR介导的Tp53基因敲除高效永生化小鼠胚胎成纤维细胞

Srisathya Srinivasan and Hsin-Yi Henry Ho

2025年01月20日 2059 阅读

Abstract

Embryonic stem cells are derived from inner cell mass of an embryo that can differentiate into every cell type in the body. Clinically, cultured red blood cell supply is of great interest. However, some of the hurdles need to be overcome. This protocol describes a two step protocol to form embryoid body and differentiate them in to hematopoietic lineage cells, especially erythroid cells.

Materials and Reagents

  1. Mouse embryonic stem cell (ES cells)
  2. 200 mM L-Glutamine (100 ml) (Life Technologies, Gibco®, catalog number: 320-5030AG )
  3. Methylcellulose (STEMCELL Technologies, catalog number: 03134 )
  4. Gelatin (STEMCELL Technologies, catalog number: 07903 )
  5. Transferrin (Human) (Boehringer-Mannheim) (Roche Diagnostics, catalog number: 652202 )
  6. MTG (Sigma-Aldrich, catalog number: M6145 )
  7. Plasma derived serum (PDS) (Animal technologies) special order from company. Typically serum contains some PDGF, which inhibit differentiation (Must call company, animal technologies, Inc)
  8. Epo (Amgen) (Thermo Fisher Scientific, catalog number: 50948385 )
  9. Protein Free Hybridoma Media II (PFHM-II) (Life Technologies, Gibco®, catalog number: 12040-077 )
  10. Ascorbic acid
  11. Penicillin/Streptomycin (Pen/Strep) (Thermo Fisher Scientific, catalog number: SV30010 )
  12. Fetal bovine serum (FBS)
  13. Fetal calf serum (FCS)
  14. Phosphate buffered saline (PBS)
  15. Trypsin
  16. EDTA
  17. NaHCO3
  18. Dulbecco's modified eagle medium (DMEM)
  19. ES-IMDM (see Recipes)
  20. Differentiation medium (see Recipes)
  21. Secondary differentiation media (see Recipes)
  22. Iscove’s modified dulbecco medium (IMDM) (Life Technologies, Gibco®, catalog number: 12200-036 , 1 L/pack) (see Recipes)
  23. Cellulase (Sigma-Aldrich, catalog number: C-1794 ) (see Recipes)

Equipment

  1. Standard table-top centrifuges
  2. T25 flask
  3. Snap cap tube
  4. Incubator
  5. Water bath
  6. Gauge needle
  7. 50 ml Falcon tube
  8. 0.22 µm filter

Procedure

English
中文翻译

文章信息

版权信息

© 2012 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Readers should cite both the Bio-protocol article and the original research article where this protocol was used:

  1. Im, H. (2012). Mouse Embryonic Stem Cell Differentiation to Hematopoietic Precursors. Bio-protocol 2(7): e144. DOI: 10.21769/BioProtoc.144.
  2. Im, H., Park, C., Feng, Q., Johnson, K. D., Kiekhaefer, C. M., Choi, K., Zhang, Y. and Bresnick, E. H. (2003). Dynamic regulation of histone H3 methylated at lysine 79 within a tissue-specific chromatin domain. J Biol Chem 278(20): 18346-18352.

    3. ris ris Download Citation in RIS Format

分类

干细胞 > 胚胎干细胞 > 维持和分化

发育生物学 > 细胞生长和命运决定

干细胞 > 成体干细胞 > 造血干细胞

您对这篇实验方法有问题吗？

在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。

0/150

tip 提问指南

+ 问题描述

写下详细的问题描述，包括所有有助于他人回答您问题的信息（例如实验过程、条件和相关图像等）。

post 发布问题
0 Q&A
pen 提交稿件