Incidence of apoptosis in clone embryos and improved development by the treatment of donor somatic cells with putative apoptosis inhibitors.
This study was conducted to promote in vitro-development of clone embryos by the treatment of donor somatic cells with hemoglobin (Hb) and/or beta-mercaptoethanol (ME), based on the analysis of apoptosis after somatic cell nuclear transfer (SCNT). Prospective, randomized study was conducted and, in vitro-matured bovine oocytes and fetal fibroblasts were provided for SCNT. In the first series of experiment, embryo apoptosis after SCNT was monitored by a terminal deoxynucleotidyl transferase-mediated d-UTP nick end-labeling assay. As results, apoptosis occurred more (P < 0.05) frequently after SCNT than after in vitro-fertilization (IVF) of control treatment. Subsequently, donor somatic cells treated with Hb (1 microg/ml) and/or ME (10 microM) were provided for SCNT. Either Hb or ME greatly reduced apoptosis (0.083 +/- 0.006 vs. 0.058-0.068 +/- 0.005), while combined treatment did not. ME was more promotive than Hb; significant increases were found in morula compaction (86%), cell numbers of blastocyst (131.3 +/- 1.3 cells/blastocyst), and inner cell mass (31.9 +/- 0.8 cells/blastocyst) cell, and the ratio of inner cell mass to trophectodermal cell numbers (0.24 +/- 0.01). In conclusion, the treatment of donor somatic cells with ME or Hb could reduce apoptosis after SCNT, resulting improved preimplantation development.
Park
E S
ES
School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea.
Hwang
W S
WS
Jang
G
G
Cho
J K
JK
Kang
S K
SK
Lee
B C
BC
Han
J Y
JY
Lim
J M
JM
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002417:Cattle; D002452:Cell Count; D019976:Cloning, Organism; D004423:Ear / Q000254:growth & development; D010064:Embryo Implantation; D004622:Embryo, Mammalian / Q000166:cytology* / Q000187:drug effects*; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro; D005347:Fibroblasts; D006454:Hemoglobins / Q000494:pharmacology*; D008623:Mercaptoethanol / Q000494:pharmacology*; D023421:Models, Animal; D011247:Pregnancy
D006454:Hemoglobins; D008623:Mercaptoethanol