a vector-based system for the differentiation of mouse embryonic stem cells toward germ-line cells
نویسندگان
چکیده
objective(s):to culture thein vitro mouse embryonic stem cells (mescs) and to direct their differentiation to germ-line cells; in present study we used a vector backbone containing the fusion construct stra8-egfp to select differentiated es cells that entered meiosis. retinoic acid was used to differentiate embryonic stem cells to germ cells. materials and methods: a fragment of stra8 gene promoter (-1400 to +7) was inserted in scai/hindiii multiple cloning site of pegfp-1 vector. theelectroporationwas done on embryonic stem cells and positive colonies were selected as puromycin-resistant after three weeks of treatment with puromycin. all-trans retinoic acid (ra) was used for differentiation of mescs at final concentration of 10-5m. the expression of protamine 1 (prm1) gene was checked as post meiotic marker in differentiated mescs after 5, 10, 15, 21 and 30 days after ra induction. results: the pcr amplification by specific primers for stra8-egfp fusion gene was detected in dna sample from mescs after electroporation and puromycin treatment. gfp-positive mesc colonies were observed after 72 hr ra induction. the protamine 1 gene was expressed after 21 days of ra induction. conclusion: in this study, we demonstrated the in vitro generation of mouse embryonic stem cells to germ cells by using a backbone vector containing the fusion gene stra8-egfp. the stra8 gene is a retinoic acid-responsive protein and is able to regulate meiotic initiation.
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A vector-based system for the differentiation of mouse embryonic stem cells toward germ-line cells
Objective(s):To culture thein vitro mouse embryonic stem cells (mESCs) and to direct their differentiation to germ-line cells; in present study we used a vector backbone containing the fusion construct Stra8-EGFP to select differentiated ES cells that entered meiosis. Retinoic acid was used to differentiate embryonic stem cells to germ cells. Materials and Methods: A fragment of Stra8 gene pr...
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عنوان ژورنال:
iranian journal of basic medical sciencesجلد ۱۷، شماره ۸، صفحات ۵۶۶-۵۷۰
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