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KMID : 0364219960390030239
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Korean Journal of Zoology
1996 Volume.39 No. 3 p.239 ~ p.247
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A Simple Embryonic Stem Cell-Based in vitro Differentiation System That Recapitulates Early Erythropoietic Events in the Mouse Embryo
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Kim Chul-Geun
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Abstract
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An embryonic stem (ES) cell-based in vitro model system was examined to determine whether a simple differentiation of embryoid bodies (EB) in the suspension medium is useful to dissect early erythropoiesis. Characteristics of the differentiating EBs were monitored for their differentiation potential to generate hematopoietic cell types by general morphology, benzidine staining and two-step colony assays, and expressivity of several erythroid marker genes by the RT-PCR analysis for total cellular RNA prepared from the differentiating EBs. Every ematopoietic lineage cells were generated from the differentiating EBs with reproducible frequencies, similar to the other sophisticated differentiation protocols. Furthermore, the globin gene switching in differentiating ES cells paralleled the sequence of events found in the mouse embryo, and such that their expression was activated by at least 12 hrs later than those of erythroid-specific transcription factors, GATA-1 and Tal-1 The erythropoietic differentiation program initiated reproducibly and efficiently in this simple differentiation system in a suspension culture, such that this system may be useful for dissection of the molecular events of early erythropoiesis.
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KEYWORD
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Gene Expression, Globin Gene, Transcription factor, Embryonic Stem Cells, in vitro Differentiation
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