isolation and enrichment of mouse female germ line stem cells
نویسندگان
چکیده
objective: the existence of female germ-line stem cells (fgscs) has been the subject of a wide range of recent studies. successful isolation and culture of fgscs could facilitate studies on regenerative medicine and infertility treatments in the near future. our aim in the present study was evaluation of the most commonly used techniques in enrichment of fgscs and in establishment of the best procedure. materials and methods: in this experimental study, after digesting neonate ovary from c57bl/6 mice, we performed 2 different isolation experiments: magnetic activated cell sorting (macs) and pre-plating. macs was applied using two different antibodies against mouse vasa homolog (mvh) and stage-specific embryonic antigen-1 (ssea1) markers. after the cells were passaged and proliferated in vitro, colony-forming cells were characterized using reverse transcription-polymerase chain reaction (rt-pcr) (for analysis of expression of oct4, nanog, c-kit, fragilis, mvh, dazl, scp3 and zp3), alkaline phosphatase (ap) activity test and immunocytochemistry. results: data showed that colonies can be seen more frequently in pre-plating technique than that in macs. using the ssea1 antibody with macs, 1.98 ± 0.49% (mean ± sdv) positive cells were yield as compared to the total cells sorted. the colonies formed after pre-plating expressed pluripotency and germ stem cell markers (oct4, nanog, c-kit, fragilis, mvh and dazl) whereas did not express zp3 and scp3 at the mrna level. immunocytochemistry in these colonies further confirmed the presence of oct4 and mvh proteins, and ap activity measured by ap-kit showed positive reaction. conclusion: we established a simple and an efficient pre-plating technique to culture and to enrich fgscs from neonatal mouse ovaries.
منابع مشابه
Isolation and Enrichment of Mouse Female Germ Line Stem Cells
OBJECTIVE The existence of female germ-line stem cells (FGSCs) has been the subject of a wide range of recent studies. Successful isolation and culture of FGSCs could facilitate studies on regenerative medicine and infertility treatments in the near future. Our aim in the present study was evaluation of the most commonly used techniques in enrichment of FGSCs and in establishment of the best pr...
متن کاملA 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...
متن کاملFunctional Concentrations of BMP4 on Differentiation of Mouse Embryonic Stem Cells to Primordial Germ Cells
متن کامل
The effect of BMP4 on mouse embryonic stem cell proliferation and differentiation into primordial germ cells
Background and Aim: Artificial gamete production from stem cells is a novel strategy for treatment of infertility. Among various stem cell sources, embryonic stem cells (ESC) can be considered as an appropriate source for in vitro formation of germ cells. In this study we evaluated the effect of BMP4 on proliferation and differentiation of mouse embryonic stem cells into primordial germ cells (...
متن کاملa 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...
متن کاملCo-culture of Mouse Embryonic Stem Cells with Sertoli Cells Promote in vitro Generation of Germ Cells
Objective(s): Sertoli cells support in vivo germ cell production; but, its exact mechanism has not been well understood. The present study was designed to analyze the effect of Sertoli cells in differentiation of mouse embryonic stem cells (mESCs) to germ cells. Materials and Methods: A fusion construct composed of a Stra8 gene promoter and the coding region of enhanced green fluorescence p...
متن کاملمنابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
cell journalجلد ۱۶، شماره ۴، صفحات ۴۰۶-۴۱۵
میزبانی شده توسط پلتفرم ابری doprax.com
copyright © 2015-2023