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Differential but beneficial effect of phytohemagglutinin on efficiency of in vitro porcine embryo production by somatic cell nuclear transfer or in vitro fertilization
Author(s) -
Gupta Mukesh Kumar,
Uhm Sang Jun,
Lee Hoon Taek
Publication year - 2007
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.20720
Subject(s) - blastocyst , biology , andrology , somatic cell nuclear transfer , embryo , zygote , in vitro , inner cell mass , apoptosis , somatic cell , embryogenesis , microbiology and biotechnology , genetics , medicine , gene
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. Mol. Reprod. Dev. 74: 1557–1567, 2007. © 2007 Wiley‐Liss, Inc.