Differential but beneficial effect of phytohemagglutinin on efficiency of in vitro porcine embryo production by somatic cell nuclear transfer or in vitro fertilization.
In vitro production of porcine embryos has become routine in most laboratories but the yield and quality of the resultant blastocysts remains sub-optimal. Phytohemagglutinin (PHA) is an N-acetylgalactosamine/galactose sugar-specific lectin with a wide variety of biological activities including mitogenesis, mediation of cell recognition and agglutination of cells. This study was therefore, designed to investigate the effect of PHA on the preimplantation development and quality of parthenogenetic, somatic cell nuclear transferred (SCNT) and in vitro fertilized (IVF) porcine embryos cultured in the absence or presence of PHA. Analysis showed that, supplementation of PHA significantly improved the blastocyst rate of parthenogenetic (70.6 +/- 0.2 vs. 51.4 +/- 0.6%) and SCNT (27.7 +/- 1.7 vs. 12.5 +/- 0.3%) embryos but not IVF embryos (25.0 +/- 14.3 vs. 20.1 +/- 12.7%). Nonetheless, PHA-treated blastocysts had higher hatching ability and contained higher cell number than control blastocysts in all the groups (P < 0.05). TUNEL labeling revealed that blastocysts cultured in the presence of PHA were less predisposed to biochemical apoptosis and showed lower indices of TUNEL, fragmentation and total apoptosis than those cultured in the absence of PHA (P < 0.05). Real time qRT-PCR analysis of parthenogenetic blastocysts revealed that PHA decreased the expression ratio of BAX:BCL-xL transcripts. Therefore, our study suggests that PHA improves the blastocyst yield and quality by enhancing blastocyst expansion, hatching and total cell number and decreasing the apoptosis. However, PHA has a differential effect on development rate of IVF derived embryos. These results may represent an approach towards achieving better preimplantation development of porcine embryos in vitro.
Gupta
Mukesh Kumar
MK
Department of Animal Biotechnology, Bio-Organ Research Center, Konkuk University, Gwangjin-Gu, Seoul, South Korea.
Uhm
Sang Jun
SJ
Lee
Hoon Taek
HT
D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D018929:Cell Culture Techniques; D002467:Cell Nucleus / Q000648:ultrastructure; D005307:Fertilization in Vitro; D053652:Nuclear Transfer Techniques; D010312:Parthenogenesis; D010835:Phytohemagglutinins / Q000494:pharmacology*; D034421:Sus scrofa / Q000196:embryology*; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism; D051020:bcl-X Protein / Q000235:genetics / Q000378:metabolism
D010835:Phytohemagglutinins; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein