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  • The Gata1low murine megakaryocyte-erythroid progenitor cells expand robustly and alter differentiation potential.

The Gata1low murine megakaryocyte-erythroid progenitor cells expand robustly and alter differentiation potential.

Biochemical and biophysical research communications (2020-05-28)
Eunju Shin, Jong-Gwan Jeong, Hyunmin Chung, Haiyoung Jung, Charny Park, Suk Ran Yoon, Tae-Don Kim, Seung Jin Lee, Inpyo Choi, Ji-Yoon Noh
초록

GATA1 is a master transcription factor of megakaryopoiesis and erythropoiesis, and loss-of-function mutation can induce accumulation of megakaryocyte-erythroid progenitors (MEPs) in mice and humans. Accordingly, the murine MEP cell line (termed G1ME2 cells) encoding doxycycline (dox)-inducible anti-Gata1 shRNA on Hprt locus has been developed. The cells were CD41+CD71+KIT+, expand under dox, stem cell factor, and thrombopoietin (TPO), and terminally differentiate into erythroid cells or megakaryocytes upon removal of dox. Surprisingly, in this study, these Gata1low murine MEPs displayed accelerated growth from around 90-100 days after cell culture, impeded megakaryocytic potential, and maintained erythropoiesis. We specified them as late G1ME2 cells and discovered that increased CD41-KIT+ population during long-term culture was the main reason for the delayed megakaryopoiesis. The CD41 expression level was partially de-repressed by PI3K/AKT inhibitors, suggesting that TPO-mediated cell survival signaling pathway might have impacted on CD41 in the late G1ME2 cells. Nevertheless, among the late cells, the CD41+KIT+ cells could still generate megakaryocytes on dox withdrawal. Taken together, G1ME2 cells could provide a good model to study molecular mechanism of hematopoiesis because of their ability to expand excessively without artificial immortalization.

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Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
Millipore
Protease Inhibitor Cocktail Set III, Animal-Free, Protease Inhibitor Cocktail Set III, Animal-Free, is a cocktail of six protease inhibitors with broad specificity for the inhibition of aspartic, cysteine & serine proteases & aminopeptidases.
Millipore
Phosphatase Inhibitor Cocktail Set III, Phosphatase Inhibitor Cocktail Set III is a ready to use cocktail of four phosphatase inhibitors for broad-spectrum inhibition of both serine/threonine and protein tyrosine phosphatases.