Unique multipotent cells in adult human mesenchymal cell populations

Abstract

We found adult human stem cells that can generate, from a single cell, cells with the characteristics of the three germ layers. The cells are stress-tolerant and can be isolated from cultured skin fibroblasts or bone marrow stromal cells, or directly from bone marrow aspirates. These cells can self-renew; form characteristic cell clusters in suspension culture that express a set of genes associated with pluripotency; and can differentiate into endodermal, ectodermal, and mesodermal cells both in vitro and in vivo. When transplanted into immunodeficient mice by local or i.v. injection, the cells integrated into damaged skin, muscle, or liver and differentiated into cytokeratin 14-, dystrophin-, or albumin-positive cells in the respective tissues. Furthermore, they can be efficiently isolated as SSEA-3(+) cells. Unlike authentic ES cells, their proliferation activity is not very high and they do not form teratomas in immunodeficient mouse testes. Thus, nontumorigenic stem cells with the ability to generate the multiple cell types of the three germ layers can be obtained through easily accessible adult human mesenchymal cells without introducing exogenous genes. These unique cells will be beneficial for cell-based therapy and biomedical research.

Novel In Vitro Culture Condition Improves the Stemness of Human Dermal Stem/Progenitor Cells

“…During in vitro culture, hDSPCs, which exhibited stem celllike characteristics in our previous study, gradually lose the expression of transcription factors associated with multipotency and self-renewal, such as SOX2 and NANOG for maintaining the characteristics of stem cells. Several previous reports suggested that the quality of the stem cells strongly depend on the culture method (Driskell et al, 2012;Grayson et al, 2007;Kuroda et al, 2010;Minchenko et al, 1994;Reiter et al, 2008;Ren et al, 2006;Rendl et al, 2005). Thus, we investigated whether hDSPCs retain their stemness in various culture conditions despite long-term culture.…”

“…For suspension cultures, collagen type IV-enriched hDSPCs were plated in HydroCell plates (Nunc) with SKP medium (Biernaskie et al, 2009;Toma et al, 2001; or 2-hydroxyethyl methacrylate (poly-HEMA) as described previously (Kuroda et al, 2010). Briefly, 15 mg of poly-HEMA was dissolved in 1 ml of 95% ethanol by shaking at 37°C; the culture dish was coated with this solution and then air-dried on the clean bench overnight.…”

The secretome of MUSE cells contains factors that may play a role in regulation of stemness, apoptosis and immunomodulation

“…Confluent cells were collected by 0.25% trypsin-EDTA, and were subjected for cell sorting to isolate Muse cells, as described previously. 8 In brief, cells were suspended in FACS Buffer, which contained 0.5% bovine serum albumin (BSA), 2 mM EDTA-2H 2 O in FluoroBrite DMEM (ThermoFisher Scientific, Japan) and were incubated with anti-human SSEA-3 antibody (1:400, BioLegend, Japan) for one hour on ice. Cells were then washed with FACS buffer for 3 times and centrifuged at 400 g for 5 min.…”

“…7 Previously, we identified unique stem cells that we named multilineage-differentiating stress enduring (Muse) cells as one to several percent of the total MSCs of the bone marrow, adipose tissue and dermis. 8 Among different cell populations in MSCs, Muse cells, which can be identified as cells positive for pluripotent surface marker SSEA-3, were initially found as stress tolerant cells, and show several aspects that may represent pluripotent-like differentiation ability of MSCs. 7,9 They are recognized to express pluripotent genes, able to differentiated into triploblastic cells from a single cells and are self-renewable, suggesting their pluripotency.…”

La biologie des cellules souchesmésenchymateuses d’origine humaine

“…De telles cellules ont été détectées dans le foie foetal, mais n'ont pas été rapportées dans la moelle osseuse. Reste l'hypothèse de la sélection de cellules médul-laires pluri potentes dans des conditions particulières de contrôle strict du degré de confluence, d'hypoxie, de sélection extensive d'un lot de sérum particulier, de conditions de stress telle une incubation prolongée dans une solution de trypsine [25,26]. De telles cellules pourraient être des vestiges de cellules souches embryonnaires ayant niché dans la moelle osseuse, ou pourraient résulter d'une reprogrammation plus ou moins complète selon un processus qui reproduirait in vivo l'induction in vitro des induced pluripotent cells.…”