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HemEx-Type9A

HemEx-TypeA is a serum-free media, which contains Polyvinyl Alcohol (PVA) and can be used for mouse Hematopoietic Stem Cells (HSCs). In general, culture media for HSCs contains serum albumin. Such albumin is required to induce cell division of HSCs. On the other hand, albumin inhibits stable undifferentiated states.

Widely used purified or recombinant albumin contains small amounts of contaminants, but are tough to remove. It is reported that such contaminants induce differentiation of HSCs. Moreover, protein's oxidation reaction induces cellular senescence. Therefore, albumin needs to be replaced by other chemicals.

Dr. Satoshi Yamazaki at The Institute of Medical Science, The University of Tokyo reported that PVA can replace albumin (1).  Moreover, PVA is a stable chemical with no oxidation reaction in medium so aging of HSCs are also suppressed. These advantages help to expand culture of HSCs while maintaining an undifferentiated state. 

Features
 

• Optimized for the expansion of mouse HSCs, Hematopoietic Stem and Progenitor Cell (HSPCs) obtained from bone marrow.
• Can expand the culture of HSCs, for more than 1 month in an undifferentiated state.
• PVA is used instead of albumin.

Protocol
 

Please see the paper of Nature, 571, 117 - 121 (2019).
Note For culture, addition of cytokine is required.

Data
 

Fig. 1 Flow cytometry analysis of HSCs
HSCs were cultured with HemEx (contains 10 ug/ml of SCF and 100 ug/ml of TPO) for 7 days in 96 well U-bottom culture plate. Lin-/Sca-1+/c-Kit+ is defined as undifferentiated mouse HSPCs in this data.

 
Reference

1. Wilkinson AC, et al. Nature, 571, 117 - 121 (2019).
2. Wilkinson AC, et al. Nat Protoc, 15 (2), 628-648 (2020).
3. Ochi K., et al. Nat Commun, 12(1): 3568. (2021).