Yoshioka Koji, Suzuki Chie, Tanaka Atsushi, Anas Idris M-K, Iwamura Shokichi
Department of Production Diseases, National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan.
Biol Reprod. 2002 Jan;66(1):112-9. doi: 10.1095/biolreprod66.1.112.
We evaluated the in vitro development of porcine zygotes that were cultured in a novel culture medium, porcine zygote medium (PZM), under different conditions and compared to in vivo development. The viability of these zygotes to full term after culture was also evaluated by embryo transfer to recipients. Porcine single-cell zygotes were collected from gilts on Day 2 after hCG injection. Culture of zygotes in PZM containing 3 mg/ml of BSA (PZM-3) produced better results in terms of proportion of Day 6 blastocysts, Day 8 hatching rate, and numbers of inner cell mass (ICM) cells and total cells in Day 8 embryos than that in North Carolina State University (NCSU)-23 medium. In culture with PZM-3, embryo development was optimized in an atmosphere of 5% CO2:5% O2:90% N2 compared to 5% CO2 in air. The ICM and total cell numbers in Day 6 embryos cultured in PZM-3 or in PZM-3 in which BSA was replaced with 3 mg/ml of polyvinyl alcohol (PZM-4) were also greater than those of NCSU-23 but less than those developed in vivo. However, no difference was found in the ratio of ICM to total cells among embryos developed in PZM-3, PZM-4, or in vivo. When the Day 6 embryos that developed in PZM-4 (99 embryos) or in vivo (100 embryos) were each transferred into six recipients, no difference was found in the farrowing rate (83.3% for both treatments) and in the number of piglets born (33 and 42 piglets, respectively). Our results indicate that porcine zygotes can develop into blastocysts in a chemically defined medium and to full term by transfer to recipients after culture.
我们评估了在新型培养基猪合子培养基(PZM)中于不同条件下培养的猪合子的体外发育情况,并与体内发育进行了比较。还通过将胚胎移植到受体中来评估这些合子培养至足月的生存能力。在注射人绒毛膜促性腺激素(hCG)后第2天从后备母猪采集猪单细胞合子。在含有3mg/ml牛血清白蛋白(BSA)的PZM(PZM-3)中培养合子,在第6天囊胚比例、第8天孵化率以及第8天胚胎的内细胞团(ICM)细胞数和总细胞数方面,比在北卡罗来纳州立大学(NCSU)-23培养基中培养产生了更好的结果。在PZM-3中培养时,与空气中5%二氧化碳的环境相比,在5%二氧化碳:5%氧气:90%氮气的气氛中胚胎发育得到优化。在PZM-3或用3mg/ml聚乙烯醇替代BSA的PZM-3(PZM-4)中培养的第6天胚胎的ICM和总细胞数也高于NCSU-23培养基中的,但低于体内发育的。然而,在PZM-3、PZM-4中或体内发育的胚胎中,ICM与总细胞的比例没有差异。当将在PZM-4中发育的第6天胚胎(99个胚胎)或体内发育的第6天胚胎(100个胚胎)分别移植到6头受体中时,产仔率(两种处理均为83.3%)和出生仔猪数(分别为33头和42头)没有差异。我们的结果表明,猪合子可以在化学成分明确的培养基中发育成囊胚,并通过培养后移植到受体中发育至足月。