翻译英文文献 Vitrification and warming of in vivo–derived porcine

SCNT

$ w' f! |$ m" D" G

3 ?! p1 ^5 f% t1 _8 L6 T  lJ. Sanchez-Osorio, C. Cuello, M.A. Gil, I. Parrilla, C. Maside,
- @% z5 H$ @! L9 Y1 tC. Almin˜ana, X. Lucas, J. Roca, J.M. Vazquez, E.A. Martinez *
: ]/ m% P9 [- u* W$ Z, [$ y- RDepartment of Animal Medicine and Surgery, Veterinary Science, University of Murcia, Murcia, Spain: i) z! l8 u* \7 h8 u1 Z
The aim of this study was to design a protocol for vitrification and warming of porcine embryos in a chemically defined medium.
6 V2 t# D0 V. Z5 Y6 M6 CA total of 663 morulae and blastocysts were collected from weaned crossbred sows (Large White-Landrace) 5 to 6 d after estrus and
0 H' c+ \- E5 Q5 |vitrified with the Superfine Open Pulled Straw method. In Experiment 1, embryos were vitrified using as a basic medium TCM-199–
0 c9 _( j/ l, ^' T- |HEPES supplemented with 20% newborn calf serum (NBCS) or with 0, 0.1%, 0.5%, or 1% polyvinyl alcohol (PVA). Nonvitrified0 b) g4 K) |/ \2 p  a. s" {
embryos were used as a fresh control group. Survival and hatching rates were evaluated after 72 h of in vitro culture to assess( ?' y5 j7 p0 p2 t* h: Q2 w! R
embryo viability. In addition, some hatched blastocysts derived from morulae and blastocysts were processed to determine the total
" d: f( t& P+ K  ^8 V' |* a! ~cell number and the cell proliferating index as measures of their quality. Within each stage of embryo development, the different( M$ u! ?/ w! z/ w, j1 }
vitrification groups and the fresh control group showed similar high embryo survival (range, 70.5  7.1% to 84.9  8.1% and
0 s6 O8 h% R8 p) j85.3  8.1% to 98.4  8.2% for morulae and blastocysts, respectively) and hatching rate (range, 46.3  10.1% to 66.7  11.2%
8 v: s  p0 f  Y/ K! Pand 73.7  11.3% to 89.4  11.2% for morulae and blastocysts, respectively) and quality after in vitro culture. In Experiment 2,1 {  Q1 Y6 @; |  K. c
embryos were vitrified using 0.1% PVA and warmed with TCM-199–HEPES–0.13 M sucrose supplemented with 20% NBCS or
9 K+ Q! @! ^/ ^either 0 or 0.1% PVA. Nonvitrified embryos were used as a fresh control group. As in Experiment 1, no significant differences were; B$ n: E1 X5 r; D
detected in embryo survival (range, 67.9  6.6% to 74.5  6.6% and 91.9  7.0% to 99.5  6.3% for morulae and blastocysts,/ E5 {4 b7 l3 i7 h$ G- |
respectively) and hatching rate (range, 47.0  7.2% to 64.8  9.9% and 89.4  7.4% to 98.2  6.9% for morulae and blastocysts," i- R3 q4 t* }( l, ?
respectively) and quality among the warming groups or among vitrified and fresh control embryos. In both experiments, the  ^" y. u4 v( c' m+ \' M% y/ u
developmental embryo stage influenced the survival and hatching rates, as well as the number of cells (P < 0.01), with the blastocyst
( s6 l. {8 k2 ]6 C3 Cstage yielding the best results. In conclusion, PVA can be used as a substitute for serum in vitrification and warming solutions5 x! I% M* T) k3 m' r) i( g& j
without detrimental effects on the in vitro development of in vivo–derived porcine morulae and blastocysts.
. k0 I7 r# ], [$ Z0 Z/ d摘要：本研究的目的是设计猪胚胎在化学确定培养液中玻璃化冷冻和复苏的程序。从发情后5-6天的杂交母猪收集663枚桑葚和囊胚，使用OPS方法进行玻璃化冷冻。试验一，使用TCM-199-HEPES作为基础液，添加20%新生牛血清（NBCS）或0、0.1%、0.5%、1% PVA对胚胎进行玻璃化冷冻。非玻璃化冷冻胚胎作为对照组。体外培养72小时后根据存活率和孵化率评估胚胎发育能力。此外，将获得的孵化囊胚进行处理通过总细胞数和细胞增殖指数评估胚胎质量。胚胎发育的不同阶段，不同玻璃化冷冻组与非冷冻组在存活率（70.5%-84.9%和85.3%-98.4%）和囊胚孵化率（46.3%-66.7%和73.7%-89.4%）方面相似。实验二，使用0.1%PVA对胚胎进行玻璃化冷冻，使用添加20%NBCS或0或0.1%PVA的TCM-199-HEPES-0.13 M蔗糖进行复苏。非玻璃化冷冻作对照。胚胎发育的不同阶段，不同玻璃化冷冻组与非冷冻组在存活率（67.9%-74.5%和91.9%-99.5%）和囊胚孵化率（47.0%-64.8%和89.4%-98.2%）方面相似。