[Developmental capacity of reconstructed embryos by transfer with peripheral blood lymphocytes in mouse].
Mouse peripheral blood lymphocytes were used as donor nuclei in nuclear transfer procedures to determine their feasibility. Lymphocytes were collected from peripheral blood by lymphocyte isolation liquid (density 1.088), and transferred into enucleated oocytes by intracytoplasmic injection. Two hours later, oocytes injected with donor cell nuclei were activated for 6 hours by treatment in modified M16 medium containing 10 mmol/L SrCl2, and then cocultured with mouse oviduct epithelial cell in mM16 medium. On early day 4 blastocysts were cultured on mouse embryonic fibroblasts in DMEM. After 4 days, ICMs ( inner cell mass) were trypsinized, seeded and cultured. Results showed that donor nuclei of lymphocyte could be reprogrammed to support the development of reconstructed embryos in vitro. The proportion of reconstructed oocytes (n = 312) that developed to 2-cell stage was 41.03%, while 9.29% and 1.92% of reconstructed oocytes developed to morula and blastocyst phase, respectively. In addition, 2 ICMs were isolated from reconstructed blastocysts, and the proportion of reconstructed oocytes developing to ICM was 0.64% (2/312). This study illustrates that it is feasible to construct embryos with peripheral blood lymphocytes in mouse, warranting further study in this area.
Cao
Hong Guo
HG
Institute of Bio-engineering, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, China.
Zhang
Yong
Y
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000502:physiology; D008214:Lymphocytes / Q000502:physiology*; D051379:Mice; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy