Comparative study of the developmental competence of cloned pig embryos derived from spermatogonial stem cells and fetal fibroblasts

Abstract

Spermatogonial stem cells (SSC) are promising resources for genetic preservation and restoration of male germ cells in humans and animals. However, no studies have used SSC as donor nuclei in pig somatic cell nuclear transfer (SCNT). This study investigated the potential for use of porcine SSC as a nuclei donor for SCNT and developmental competence of SSC‐derived cloned embryos. In addition, demecolcine was investigated to determine whether it could prevent rupture of SSC during SCNT. When the potential of SSC to support embryonic development after SCNT was compared with that of foetal fibroblasts (FF), SSC‐derived SCNT embryos showed a higher (p < .05) developmental competence to the blastocyst stage (47.8%) than FF‐derived embryos (25.6%). However, when SSC were used as donor nuclei in the SCNT process, cell fusion rates were lower (p < .05) than when FF were used (61.9% vs. 75.8%). Treatment of SSC with demecolcine significantly (p < .05) decreased rupture of SSC during the SCNT procedure (7.5% vs. 18.8%) and increased fusion of cell‐oocyte couplets compared with no treatment (74.6% vs. 61.6%). In addition, SSC‐derived SCNT embryos showed higher blastocyst formation (48.4%) than FF‐derived embryos without (28.4%) and with demecolcine treatment (17.4%), even after demecolcine treatment. Our results demonstrate that porcine SSC are a desirable donor cell type for production of SCNT pig embryos and that demecolcine increases production efficiency of cloned embryos by inhibiting rupture of nuclei donor SSC during SCNT.

Ex Situ Conservation and Genetic Rescue of Endangered Polish Cattle and Pig Breeds with the Aid of Modern Reproductive Biotechnology – A Review

“…To sum up, the efficiency of intra-species and inter-species somatic cell cloning of domesticated mammals is affected, to a great extent, by three factors: the provenance of frozen/thawed and ex vivo-expanded nuclear donor cell lines (NDCLs) that provide genetic resources from different livestock species or breeds (Samiec and Skrzyszowska, 2010;Li et al, 2013;Olivera et al, 2016;Lee et al, 2019), the morphological, cytogenetic, cytophysiological and molecular quality parameters of the NDCLs (Samiec et al, 2013(Samiec et al, , 2019Zhang et al, 2019;Wiater et al, 2021); and the subsequent incidence of programmed (apoptotic or autophagic) cell death in the development of cloned embryos (Chi et al, 2017;Samiec et al 2015;Jeong et al, 2020). This efficiency is also dependent on such molecular mechanisms as the epigenetic ability of the nuclear donor cell-inherited genome to be reprogrammed in SCNT-derived oocytes and result in cloned embryos (Samiec and Skrzyszowska, 2018;Deng et al, 2020;Sampaio et al, 2020;Wang et al, 2020b) and the intergenomic crosstalk between nuclear and mitochondrial DNA fractions in the host cytoplasm of nuclear recipient oocytes and the blastomeres of the developing cloned embryos (Srirattana et al, 2011;Samiec and Skrzyszowska, 2021;Takeda, 2019;Xu et al, 2019;Magalhães et al, 2020) (Figure 1).…”

Crosstalk between Peroxisomal Activities and Nrf2 Signaling in Porcine Embryos

“…The production of transgenic pigs and xenotransplantation are pivotal in translational biomedical research [ 1 , 2 ], and the development of optimal procedures for embryo production are essential [ 3 ]. However, overcoming the four-cell block and maximizing the developmental potentials of porcine embryos still remains an unsolvable assignment for researchers [ 4 , 5 ]; therefore, attempts to improve the porcine IVC system and transgenic pig production using either chemical treatment or assistive reproductive technologies (ARTs) have been continuously reported [ 6 , 7 , 8 , 9 ]. Interestingly, both phytanic acid (PA) (a branched fatty acid (FA) and melatonin (a mammalian pineal gland- and peripheral reproductive organ-derived natural hormone) are suggested to be involved in lipid metabolism in mammalian cells [ 10 , 11 ], oocytes [ 12 , 13 , 14 ], and embryos [ 7 , 15 , 16 ].…”

Genome stabilization by RAD51‐stimulatory compound 1 enhances efficiency of somatic cell nuclear transfer‐mediated reprogramming and full‐term development of cloned mouse embryos

“…3 Various efforts have been undertaken to comprehensively recognize the pivotal molecular factors affecting successful generation of mammalian SCNT-cloned embryos, conceptuses and offspring. These factors determine among others (a) the provenance of nuclear donor cells, [4][5][6][7] (b) quality of nuclear recipient oocytes related to their meiotic, epigenomic and cytoplasmic maturity status. [8][9][10] The aforementioned factors also determine (c) epigenetic reprogrammability of donor cell nuclear genome in SCNT-derived oocytes and resultant embryos, 11-15 (d) intergenomic crosstalk between nuclear and mitochondrial compartments in SCNT-derived oocytes and cloned embryos [16][17][18][19][20] and (e) the incidence of apoptosis-or autophagydependent events in the ex vivo-expanded nuclear donor cells and cultured cloned embryos.…”