Isolation and Characterization of SSEA3⁺ Stem Cells Derived from Goat Skin Fibroblasts

Abstract

Novel stem cells expressing stage-specific embryonic antigen 3 (SSEA-3) reside among human dermal fibroblasts and are known as multilineage-differentiating stress-enduring (Muse) cells. They enhance the generation efficiency of induced pluripotent stem cells. However, Muse cells have only been found in humans. We aimed to isolate SSEA3-positive cells from terminally differentiated skin fibroblasts of adult goat and determine their pluripotency. Cell clusters from SSEA3(+) populations possessed stem cell-like morphological features and normal karyotypes, were consistently positive for alkaline phosphatase, and expressed stem cell pluripotency markers. These SSEA3(+) cells remained undifferentiated over eight passages in suspension culture and were able to differentiate into cells of all three germ layers in vitro and in vivo. Our combined findings suggest that a subset of adult stem cells expressing SSEA3 also exist among adult goat skin fibroblasts. We are the first to report that multipotent adult goat cells exist among terminally differentiated goat skin in suspension culture. Our results also provide a promising platform for generation of a transgenic goat, because the undifferentiated state of stem cells was thought to be more efficient as donor cells for somatic cell nuclear transfer.

Identification of SSEA-1 expressing enhanced reprogramming (SEER) cells in porcine embryonic fibroblasts

“…11,12 Muse cells are sorted based on co-expression of SSEA-3 and CD105 and possess at least some iPSC characteristics, such as increased expression of pluripotency markers, self-renewal, and multipotential differentiation capabilities. 10 A similar cell population has also been reported in skin fibroblasts derived from goat, 13 suggesting that the cell type is not confined to a single species. These findings imply that the cellular expression profile of the original cell type might play an important role in the reprogramming process.…”

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

“…It was a major challenge to maintain the undifferentiated state of SSEA-4 + cells because that little is known about culture conditions that support SSEA-4 + cells proliferation in vitro. Previous reports have shown that stem cells were isolated from skin tissue via selective growth of floating spheres [ 7 , 23 , 25 , 27 ]. Therefore, we maintained SSEA-4 + cells in suspension culture using optimized medium containing EGF and bFGF in order to retain the ability of cells to self-renew, as described in a previous study [ 25 ].…”

“…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.…”

Pluripotent nontumorigenic multilineage differentiating stress enduring cells (Muse cells): a seven-year retrospective

“…M clusters express pluripotent markers such as NANOG, Oct3/4, Par-4, Sox2, and TRA1-60 in addition to markers from mesodermal (NK2-5), endodermal (GATAφ, α-fetoprotein), and ectodermal (MAP2) cell lines spontaneously and under media-specific induction [ 20 ]. Seven independent groups have confirmed this inherent pluripotent phenotype over the last 7 years [ 14 , 17 , 18 , 21 , 22 , 27 , 28 ]. These groups have found Muse cells throughout different mesenchymal tissues including bone marrow, skin, and adipose tissue retaining the same pluripotent potential and Muse cell phenotype [ 14 , 17 , 18 ].…”