Developmental Potential of Cloned Goat Embryos from an SSEA3⁺ Subpopulation of Skin Fibroblasts

Abstract

Previous studies have demonstrated that skin stem cells expressing the pluripotency marker stage-specific embryonic antigen 3 (SSEA3) are easier to reprogram into induced pluripotent stem cells (iPSCs) than skin fibroblasts. Furthermore, it is widely speculated that the undifferentiated state may make stem cells more efficient donor cells for somatic cell nuclear transfer (SCNT). In this study, we isolated SSEA3(+) cells from goat skin fibroblast cells (SFCs) using fluorescence-activated cell sorting (FACS) and examined expression of pluripotency markers and in vitro development of cloned embryos following SCNT. Results showed that cell clusters from SSEA3(+) cells were consistently positive for alkaline phosphatase staining and pluripotency markers, Nanog, Oct4, Sox2, and SSEA3. The cleavage rate of cloned embryos derived from SSEA3(+) cells did not differ compared with SFCs (70.5±0.8% and 68.4±2.1%, respectively), but was significantly higher compared with SSEA3(-) cells (64.9±1.6%, p<0.05). The blastocyst rate was significantly increased in the SSEA3(+) cell group compared with the SFC and SSEA3(-) cell groups (30.3±1.2% vs. 21.2±0.9 and 19.0±1.0%, respectively, p<0.05). The quality of cloned blastocysts from SSEA3(+) cells was higher compared with SFCs and SSEA3(-) cells, based on total cell number and number of apoptotic cells per blastocyst. These findings suggest that using SSEA3(+) cells as donors for SCNT is beneficial for enhancing in vitro development and quality of cloned goat embryos.

Application of a Novel Population of Multipotent Stem Cells Derived from Skin Fibroblasts as Donor Cells in Bovine SCNT

“…These porcine skin stem cells also had the intrinsic ability to differentiate into oocyte-like cells [ 26 ]. Furthermore, a subpopulation of multipotent stem cells derived from goat skin enhanced the development and quality of cloned goat embryos in vitro [ 27 , 28 ]. While these studies demonstrate the potential of these multipotent stem cells in improving cloning strategies, further work needs to be done to fully characterize these cells in order to maximize cloning efficacy and to streamline the process of transgenic animal production.…”

A mystery unraveled: nontumorigenic pluripotent stem cells in human adult tissues

“…Liu et al . isolated SSEA+ Muse cells from goat skin fibroblast cells (SFC) by FACS in order to investigate their application for somatic cell nuclear transfer (SCNT) [20]. They demonstrated that 2 – 4% of SFCs are SSEA+ Muse cells and that when applied as donor cells for SCNT, Muse cells contributed to a higher blastocyst rate than SSEA- SFCs in cloned embryos.…”

“…They demonstrated that 2 – 4% of SFCs are SSEA+ Muse cells and that when applied as donor cells for SCNT, Muse cells contributed to a higher blastocyst rate than SSEA- SFCs in cloned embryos. Furthermore, blastocysts from donor Muse cells demonstrated enhanced quality as determined by total cell number and rate of apoptosis [20]. Confirming the existence of Muse cells in other mammalian species is a major step in the investigation of their potential and will undoubtedly inspire further animal studies to demonstrate their applicability for stem cell therapies.…”

“…Multilineage Differentiating Stress Enduring (Muse) cells were isolated under severe cellular stress conditions, from human bone marrow and dermal fibroblasts retaining self-renewing properties despite their lack of teratoma formation when injected into immune-deficient mice [17]. Muse cells have since been isolated from human adipose tissue lipoaspirates [18], commercially available adipose stem cells (ASCs) [19] and goat fibroblasts [20]. Amongst a plethora of attractive qualities demonstrated by Muse cells, controlled proliferation in the absence of teratoma formation sets them apart from ES and iPS cells and offers an innovative and exciting avenue of exploration for the potential applications of these pluripotent stem cells to treat various human diseases.…”

Deriva de células epiteliales de tejido de piel descongelado de Ovis canadensis mexicana para la formación de un banco de germoplasma

“…La formación de bancos de germoplasma de células somáticas en especies silvestres, requiere de la colecta de tejidos de animales vivos, generalmente en cautiverio, de los que solo pueden obtenerse tejidos de muy pequeñas dimensiones, generalmente piel, para evitar dañarlos. Las células somáticas podrán entonces ser utilizadas en la producción de embriones por transferencia nuclear, que utiliza fibroblastos derivados de diversos tejidos como la piel y las células de la granulosa, entre otras (Folch et al 2009;León-Quinto et al 2011;Kurd et al 2013;Liu et al 2013).…”