Establishment and characterization of porcin trophoblast stem cell lines from somatic cell nuclear-transferred blastocysts

Abstract

Abstract Background: Genetically modified pigs are considered ideal models for studying human diseases and potential sources for xenotransplantation research. However, the somatic cell nuclear transfer (SCNT) technique utilized to generate these cloned pig models has low efficiency and fetuses exhibit limited developmental potential for abnormalities in the placenta. Methods: Herein, we report, for the first time, the establishment of porcine trophoblast stem cell (TSC) lines using SCNT and in vitro fertilized (IVF) blastocysts by activation of Wing-less/Integrated (Wnt) and epidermal growth factor (EGF) pathways, inhibition of transforming growth factor-β (TGFβ) and Rho-associated protein kinase (ROCK) pathways, and supplementation with ascorbic acid. We also compared the transcripts of TSC lines originating from SCNT and IVF embryos and their differentiated lineages. Results: A total of 19 porcine TSC lines were established from SCNT and IVF blastocysts (TSCNT and TSCIVF lines) showing typical TSC characteristics. Compared with the TSCIVF lines, TSCNT lines showed distinct expression patterns suggesting unique TSCNT characteristics, including decreased mRNA expression levels of genes related to apposition, steroid hormone biosynthesis, angiopoiesis, and RNA stability. Conclusions: This study provides valuable information and a powerful model for studying the abnormal development and dysfunction of trophoblasts and placentas in cloned pigs.