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Establishment of erythroleukemic GAK 14 cells and characterization of GATA 1 N ‐terminal domain
Author(s) -
Mukai Harumi Y.,
Suzuki Mikiko,
Nagano Masumi,
Ohmori Shin'ya,
Otsuki Akihito,
Tsuchida Kouhei,
Moriguchi Takashi,
Ohneda Kinuko,
Shimizu Ritsuko,
Ohneda Osamu,
Yamamoto Masayuki
Publication year - 2013
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/gtc.12084
Subject(s) - gata1 , biology , haematopoiesis , stromal cell , microbiology and biotechnology , gene knockdown , erythropoiesis , cell culture , stem cell , cancer research , medicine , genetics , anemia
GATA 1 is a transcription factor essential for erythropoiesis and megakaryopoiesis. It has been found that Gata1 gene knockdown heterozygous female ( Gata1 G1.05/+ ) mice spontaneously develop erythroblastic leukemias. In this study, we have generated a novel Gata1 knockdown erythroblastic cell line, designated GAK 14, from the leukemia cells in the Gata1 G1.05/+ mice. Although GAK14 cells maintain immature phenotype on OP 9 stromal cells in the presence of erythropoietin and stem cell factor, the cells produce G r‐1‐, M ac1‐, B 220‐, CD 3e‐ or CD 49b‐positive hematopoietic cells when co‐cultured with DAS 104‐8 feeder cells. However, GAK 14 cells did not produce erythroid and megakaryocytic lineages, perhaps due to the absence of GATA 1. Indeed, GAK 14 cells became capable of differentiating into mature erythroid cells when complemented with full‐length GATA 1 and co‐cultured with fetal liver–derived FLS 5 stromal cells. This differentiation potential was impaired when GATA 1 lacking the N ‐terminal domain was complemented. The N ‐terminal domain is known to contribute to the pathogenesis of transient abnormal myelopoiesis and acute megakaryoblastic leukemia related to D own syndrome. These results thus showed that GAK 14 cells will serve as a powerful tool for dissecting domain function of GATA 1 and that the GATA 1 N ‐terminal domain is essential for the erythroid differentiation of GAK 14 cells.