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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: 15465

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The authors used this protocol in:

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

May 2003

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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
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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

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