CZB和M16对胚胎发育的影响.pdfVIP

MOLECULAR REPRODUCTION AND DEVELOPMENT 66:126–133 (2003) Effects of Donor Oocytes and Culture Conditions on Development of Cloned Mice Embryos SHAORONG GAO, MICHELLE McGARRY, HELEN PRIDDLE, TRICIA FERRIER, BIANCA GASPARRINI, JUDY FLETCHER, LINDA HARKNESS, PAUL DE SOUSA, JIM McWHIR, AND IAN WILMUT* Department of Gene Expression and Development, The Roslin Institute, Roslin, Scotland, United Kingdom ABSTRACT Mice have been successfully nuclear transfer embryos developed into live offspring cloned from somatic and embryonic stem (ES) cells (Wilmut et al., 1997; Kato et al., 1998; Wakayama et al., using the ‘‘Honolulu method.’’ In the present study, 1998; Baguisi et al., 1999; Onishi et al., 2000; Polejaeva different donor oocytes and different culture conditions et al., 2000). The reason for this poor survival rate is were compared to evaluate the developmental poten- unknown to date. tial of nuclear transfer embryos reconstructed with an In most mouse cloning studies, B6D2F1 (C57BL/ inbred ES cell line HM-1. Oocytes were recovered from 6 DBA/2) or B6C3F1 (C57BL/6 C3H/He) female mice two different F1 donors B6D2F1 (C57BL/6 DBA/2) were used as oocyte donors for nuclear transfer and CZB and B6CBAF1 (C57BL/6 CBA). There was no effect medium was employed to culture the cloned embryos of oocyte origin on development of cloned embryos to under 5% CO2 in air to the morulae/blastocyst stage. the morulae/blastocyst stage (B6D2F1 44.1% vs. So it is a question whether these are the ideal condi- B6CBAF1 45.0%), and the transferred embryos could tions for mouse nuclear transfer and if it is possible to develop to term. Two culture conditions were com- improve nuclear transfer efficiency by changing