Wang Ying, Ren Jilong, Song Yuran, Hai Tang, Zhou Qi, Liu Zhonghua
College of Life Science, Northeast Agricultural University of China, Harbin 150030, Heilongjiang, China.
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Sheng Wu Gong Cheng Xue Bao. 2016 Jul 25;32(7):975-985. doi: 10.13345/j.cjb.150433.
With the advancements of stem cells and regenerative medicine, interspecies chimera has become a hot topic and will pave a new way of providing donor sources in organ transplantation. However, the interspecies chimera is confronted with a number of scientific questions and technical obstacles, including selections of appropriate embryonic stage and appropriate culture medium; those factors will deeply influence the developmental balance between donor cells and receptor embryos. Due to its relatively rapid reproductive cycle and similar organ size to human's, porcine is a very potential donor candidate to study these questions. To compare the development and chimeric efficiency of interspecies embryos, we tested and evaluated three different culture systems, PZM-3 (Porcine zygotic medium), culture medium for iPSCs (N2B27) and 3.5 h of N2B27 before PZM-3 (N2B27(3.5 h)), and two different embryonic stages, 8-cell and blastocyst in mouse-porcine chimeric embryos using parthenogenetically activated porcine embryos and mouse induced pluripotent stem cells (miPS). The results showed that, PZM-3 was beneficial for both development of chimeric embryos and miPSCs proliferation in porcine embryos in the 8-cell injection group. After early blastocyst injection, the chimeric efficiency did not appear significantly different among the three culture systems but was lower than 8-cell injection. In summary, the results suggest that 8-cell injection and PZM-3 culture medium are more beneficial to the in vitro development and chimeric efficiency of mouse-porcine chimeric embryos.
随着干细胞和再生医学的发展,种间嵌合体已成为一个热门话题,并将为器官移植提供供体来源开辟一条新途径。然而,种间嵌合体面临着许多科学问题和技术障碍,包括合适胚胎阶段和合适培养基的选择;这些因素将深刻影响供体细胞与受体胚胎之间的发育平衡。由于猪的繁殖周期相对较快且器官大小与人类相似,猪是研究这些问题的极具潜力的供体候选者。为了比较种间胚胎的发育和嵌合效率,我们测试并评估了三种不同的培养系统,即猪合子培养基(PZM - 3)、诱导多能干细胞培养基(N2B2)以及在PZM - 3之前使用3.5小时的N2B2(N2B2(3.5 h)),以及两个不同的胚胎阶段,即使用孤雌激活的猪胚胎和小鼠诱导多能干细胞(miPS)构建的小鼠 - 猪嵌合胚胎中的8细胞期和囊胚期。结果表明,在8细胞注射组中,PZM - 3有利于嵌合胚胎的发育以及猪胚胎中miPSCs的增殖。在早期囊胚注射后,三种培养系统之间的嵌合效率没有显著差异,但低于8细胞注射。总之,结果表明8细胞注射和PZM - 3培养基对小鼠 - 猪嵌合胚胎的体外发育和嵌合效率更有利。