| Publication number |
CLPUB00425 |
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| Authors |
Walasek M.A., Kardel M., Noort R., Chen J., Chau C., Yu I., Chang W., Wognum B., Szilvassy S., Thomas T., Eaves A., Louis S. |
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| Title |
STEMdiff hematopoietic kit reproducibly generates functional hematopoietic progenitor cells from human pluripotent stem cells. |
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| Citation |
Exp. Hematol. 53 Suppl. 1:S59-S59(2017) |
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| Web pages |
https://exphem-53-s1.elsevierdigitaledition.com/files/assets/common/downloads_cc0dbb4c/publication.pdf |
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| Abstract |
Hematopoietic cells generated from human pluripotent stem cells (hPSCs)
can be used to model blood diseases and as an alternate source of blood
cells for transplantation. However, robust methods to differentiate hPSCs
to hematopoietic progenitor cells (HPCs) have been difficult to develop.
The STEMdiff Hematopoietic Kit reproducibly generates HPCs from multiple
embryonic (H1, H9) and induced pluripotent (WLS-1C, STiPS-F016, STiPS-M001,
STiPS-B004) stem cell lines under serum-and feeder-free conditions.
Briefly, hPSC aggregates were plated on Corning Matrigel in TeSR medium.
The cells were then sequentially incubated in two STEMdiff differentiation
media and harvested on day 10 or 12. Most HPCs were detected in the
supernatant fraction of the cultures at a frequency of 41 +- 2% CD34+CD45+
HPCs (mean +- SEM, n=69, day 12), with an average yield of 4.6x10^5 +- 0.41
HPCs per well of a 12-well plate. Multilineage differentiation potential
of the cells was tested in HPC culture assays. The frequency of
hematopoietic colony-forming unit (CFU) was assessed in serum-free
MethoCult medium (H4636) designed specifically to support balanced
erythroid and myeloid colony growth from hPSC-derived HPCs. All tested
cell lines showed high CFU frequencies of 441 +- 116 CFU per 10^4 day 10
HPCs (n=6) and 85 +- 10 CFU/10^4 per day 12 HPCs (n=6). Interestingly, the
distribution of myeloid and erythroid colonies was different for day 10
and day 12 cells. Most colonies derived from day 10 HPCs were erythroid
(65% +- 5), whereas myeloid colonies (73% +- 9) were the main subtype
observed in day 12 cells. The same trend was observed for all tested cell
lines. Erythroid potential of hPSC-derived HPCs was further tested in 14
day liquid expansion cultures using StemSpan SFEM II medium with Erythroid
Expansion Supplement. As a proof-of-principle, four hPSC lines were tested
and showed 8- to 56-fold expansion of total cell number and high purity of
79-95% CD71+GlyA+ erythroid cells. In summary, the STEMdiff Hematopoietic
Kit reproducibly differentiates hPSCs to functional HPCs expressing key
cell surface markers and capable of multilineage differentiation.
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| Cell lines |
CVCL_9S49; SCTi001-A CVCL_9S50; SCTi002-A CVCL_VF00; STiPS-B004 |
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