| dc.description.abstract | Female germline stem cells (FGSCs) have been successfully established
and differentiated into oocytes in small animals. However, whether FGSCs can be
established and differentiated to oocyte-like cells (OLCs) in large animals like
bovine is still unclear. Therefore, the aim of this research is to establish FGSCs
from bovine ovaries and differentiate them to OLCs. In this experiment, cortical
tissues were cut from the ovarian surface epithelium of bovine ovaries to isolate
FGSCs. The isolated FGSCs were characterized with germ cell marker DAZL and
continued to exam the differentiation potential to oocyte-like cells by co-culture
with granulosa cells isolated from pre-antral bovine follicles. The growth factors
such as estradiol, androgen and follicle-stimulating hormone (FSH) also being
tested and applied for culture medium of FGSCs via in vitro growth of small
oocytes. Furthermore, primordial oocytes also be collected from the bovine ovaries
as a control group to compare with OLCs by immunostaining with germ cell marker
VASA and oocyte marker GDF9. As a result, the FGSCs isolated from 1-week
culture expressed DAZL marker and the enlarged cells (up to 20-30μm in
diameter) picked up from 7-day co-culture dish expressed both VASA and GDF9
in their cytoplasm. In addition, the co-culture with granulosa cells also shows
positive result in promoting the differentiation to OLCs comparing with mouse
embryonic fetus (MEF) feeder layer. In conclusion, this research have successfully
isolated and characterized bovine FGSCs but the differentiation to OLCs still need
more improvement. We suggested that the co-culture with granulosa cells can
enhance the FGSCs differentiation to OLCs.
Keyword: FGSCs, OLCs, maintenance, differentiation, oogenesis, granulosa cells,
immunostaining. | en_US |
