韵母风情:求翻译~~

Materials and methods
Reagents
All reagents were purchased from Sigma unless stated otherwise.
Mice
Mice were purchased from Taconic (C57Bl/6J X C3H/HeN, referred
to as B6C3) or Charles River (albino ICR). Oct4–GFP homozygous
transgenic mice (OG2), described previously
(Yoshimizu et al. 1999; Szabo et al. 2002), were kindly provided
by J.R. Mann (Beckman Research Institute of the City of Hope,
Duarte, CA). Female B6C3 mice, four- to six-week-old, were
superovulated with 7.5 U of pregnant mare’s serum gonadotropin
(PMSG), followed by 7.5 U of human chorionic gonadotropin
(hCG) 48 h later, to provide the recipient oocytes for nuclear
transfer. The nucleus-donor cells were obtained from
ICR × OG2 mice (OG2F1) and were therefore Oct4–GFP+/?. Female
ICR mice were used as recipients for embryo transfer.
Animals were maintained and used for experimentation according
to the guidelines of the Institutional Animal Care and Use
Committee of the University of Pennsylvania.
Recipient oocytes for nuclear transfer
Ovulated cumulus–oocyte complexes were collected 14 h after
hCG injection of PMSG-primed B6C3 females. Cumulus cells
were removed by hyaluronidase treatment (50 U/mL in HEPESbuffered
CZB medium at 20°C). Cumulus-free oocytes were
washed free of hyaluronidase and incubated in M16 medium
(see Embryo Culture).
Nucleus-donor cells
Oct4–GFP transgenic donor cells were used to visualize (re-)
expression of the transgene after nuclear transfer. Adult cumulus
cells were isolated from the cumulus–oocyte complexes
ovulated by OG2F1 females as described above. Fetal germ cells
were mechanically isolated from the gonads of 13.5–16.5-dpc
OG2F1 male fetuses and used within 3 h. The identity of germ
cells was ascertained by morphology and Oct4–GFP expression,
germ cells being the only GFP-positive cells after gastrulation
(Yoshimizu et al. 1999). Mouse embryonic fibroblasts carrying
the CD8–GFP transgene (unknown genomic background) were
obtained from U. von Andrian (The Center for Blood Research,
Harvard Medical School, Boston, MA). The nucleus-donor cells
were washed in HEPES-CZB medium, centrifuged, and suspended
in the micromanipulation medium (see below).
请大家帮帮忙，翻译一下这段论文,谢谢~!!
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