Developmental expression of lineage specific genes in porcine embryos of different origins.
OBJECTIVE
This study compared the expression of genes involved in pluripotency, segregation of inner cell mass (ICM) and trophectoderm (TE), and primitive endoderm (PE) formation in porcine embryos produced by in vitro fertilization (IVF), parthenogenetic activation (PA), and nuclear transfer (NT) using either fetal fibroblasts (FF-NT) or mesenchymal stem cells (MSC-NT).


METHODS
Blastocyst formation and total cell number were analyzed. The expression patterns of transcripts, including SRY-related HMG-box gene 2 (SOX2), reduced expression gene 1 (REX1/ZFP42), LIN28, caudal type homeobox 2 (CDX2), TEA domain family member 4 (TEAD4), integrin beta 1 (ITGB1) and GATA6 were assessed at the 4-8 cell and blastocyst stage embryos by real-time PCR.


RESULTS
Developmental rates to blastocyst stage and total cell number were higher in IVF and PA embryos than in NT embryos. But MSC-NT embryos had increased blastocyst formation and higher total cell number compared to FF-NT embryos. The relative expressions of transcripts were higher in blastocysts than in 4-8 cell stage embryos. The mRNA expression levels of SOX2 and REX1 were largely similar in embryos of different origins. However, the genes such as LIN28, CDX2, TEAD4, ITGB1 and GATA6 showed the differential expression pattern in PA and NT embryos compared to IVF embryos. Importantly, the transcript levels in MSC-NT embryos were relatively less variable to IVF than those in FF-NT embryos.


CONCLUSIONS
MSCs seem to be better donors for porcine NT as they improved the developmental competency, and influenced the expression pattern of genes quite similar with IVF embryos than that of FFs.
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
B Mohana
BM
Kumar
Geun Ho
GH
Maeng
Ryoung Hoon
RH
Jeon
Yeon Mi
YM
Lee
Won Jae
WJ
Lee
Byeong Gyun
BG
Jeon
Sun A
SA
Ock
Gyu Jin
GJ
Rho
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D019070:Cell Lineage; D004475:Ectoderm / Q000166:cytology / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D047108:Embryonic Development; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D005307:Fertilization in Vitro / Q000379:methods; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D050993:GATA6 Transcription Factor / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D010312:Parthenogenesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013552:Swine; D014158:Transcription, Genetic
D015415:Biomarkers; D050993:GATA6 Transcription Factor; D012333:RNA, Messenger; D055748:SOXB1 Transcription Factors