Barberi T, Willis LM, Socci ND, Studer LDerivation of multipotent mesenchymal precursors from human embryonic stem cells. PLoS Med 2:e161

Memorial Sloan-Kettering Cancer Center, New York, New York, United States
PLoS Medicine (Impact Factor: 14.43). 07/2005; 2(6):e161. DOI: 10.1371/journal.pmed.0020161
Source: PubMed


Human embryonic stem cells provide access to the earliest stages of human development and may serve as a source of specialized cells for regenerative medicine. Thus, it becomes crucial to develop protocols for the directed differentiation of embryonic stem cells into tissue-restricted precursors.
Here, we present culture conditions for the derivation of unlimited numbers of pure mesenchymal precursors from human embryonic stem cells and demonstrate multilineage differentiation into fat, cartilage, bone, and skeletal muscle cells.
Our findings will help to elucidate the mechanism of mesoderm specification during embryonic stem cell differentiation and provide a platform to efficiently generate specialized human mesenchymal cell types for future clinical applications.

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Available from: Tiziano Barberi, Mar 20, 2015
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    • "GFP and dystrophinpositive fibers were detected in the muscles of only two out of eight mice transplated with cells, and one of these mice had as few as 30 of these fibers (Bhagavati and Xu 2005). Together, these results indicate that the co-culture system is not efficient in the induction of myogenic differentiation of PSCs (Barberi et al. 2005; Bhagavati and Xu 2005; Archacka et al. 2014). In addition, although the co-culture system yields important information about myogenic potential of stem cells and reciprocal interactions between co-cultured cells, this method is not likely to be clinically suitable. "
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    ABSTRACT: Pluripotent stem cells (PSCs), such as embryonic stem cells or induced pluripotent stem cells are a promising source of cells for regenerative medicine as they can differentiate into all cell types building a mammalian body. However, protocols leading to efficient and safe in vitro generation of desired cell types must be perfected before PSCs can be used in cell therapies or tissue engineering. In vivo, i.e. in developing mouse embryo or teratoma, PSCs can differentiate into skeletal muscle, but in vitro their spontaneous differentiation into myogenic cells is inefficient. Numerous attempts have been undertaken to enhance this process. Many of them involved mimicking the interactions occurring during embryonic myogenesis. The key regulators of embryonic myogenesis, such as Wnts proteins, fibroblast growth factor 2, and retinoic acid, have been tested to improve the frequency of in vitro myogenic differentiation of PSCs. This review summarizes the current state of the art, comparing spontaneous and directed myogenic differentiation of PSCs as well as the protocols developed this far to facilitate this process.
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    • "Barbet et al. identified differentially expressed genes utilising a Taqman low-density array and reported that hESC-MSC clustered more closely with BM-hMSC than with undifferentiated hESC (Barbet et al., 2011). Barberi et al. utilised affymetrix oligonucleotide arrays on a CD73 + sorted population derived from hESC and compared these with BM-hMSC (Barberi et al., 2005). Similar to our data the authors found enrichment for genes associated with vascular development and inflammatory response/wound healing. "
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    ABSTRACT: Human bone marrow-derived stromal (skeletal) stem cells (BM-hMSC) are being employed in an increasing number of clinical trials for tissue regeneration. A limiting factor for their clinical use is the inability to obtain sufficient cell numbers. Human embryonic stem cells (hESC) can provide an unlimited source of clinical grade cells for therapy. We have generated MSC-like cells from hESC (called here hESC-stromal) that exhibit surface markers and differentiate to osteoblasts and adipocytes, similar to BM-hMSC. In the present study, we used microarray analysis to compare the molecular phenotype of hESC-stromal and immortalised BM-hMSC cells (hMSC-TERT). Of the 7379 genes expressed above baseline, only 9.3% of genes were differentially expressed between undiffer-entiated hESC-stromal and BM-hMSC. Following ex vivo osteoblast induction, 665 and 695 genes exhibited ≥2-fold change (FC) in hESC-stromal and BM-hMSC, respectively with 172 genes common to both cell types. Functional annotation of significantly changing genes revealed similarities in gene ontology between the two cell types. Interestingly, genes in categories of cell adhesion/motility and epithelial–mesenchymal transition (EMT) were highly enriched in hESC-stromal whereas genes associated with cell cycle processes were enriched in hMSC-TERT. This data suggests that while hESC-stromal cells exhibit a similar molecular phenotype to hMSC-TERT, differences exist that can be explained by ontological differences between these two cell types. hESC-stromal cells can thus be considered as a possible alternative candidate cells for hMSC, to be employed in regen-erative medicine protocols.
    Full-text · Article · Jul 2015
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    • "Several groups described strategies to derive MSC-like cells from either ESCs (Barberi et al., 2005; Boyd et al., 2009) or iPSCs (Liu et al., 2012; Diederichs and Tuan, 2014; Zhang et al., 2011). These approaches were based on coculture with primary MSCs, growth factor combinations , or spontaneous differentiation in embryoid bodies (EBs). "
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    ABSTRACT: Standardization of mesenchymal stromal cells (MSCs) remains a major obstacle in regenerative medicine. Starting material and culture expansion affect cell preparations and render comparison between studies difficult. In contrast, induced pluripotent stem cells (iPSCs) assimilate toward a ground state and may therefore give rise to more standardized cell preparations. We reprogrammed MSCs into iPSCs, which were subsequently redifferentiated toward MSCs. These iPS-MSCs revealed similar morphology, immunophenotype, in vitro differentiation potential, and gene expression profiles as primary MSCs. However, iPS-MSCs were impaired in suppressing T cell proliferation. DNA methylation (DNAm) profiles of iPSCs maintained donor-specific characteristics, whereas tissue-specific, senescence-associated, and age-related DNAm patterns were erased during reprogramming. iPS-MSCs reacquired senescence-associated DNAm during culture expansion, but they remained rejuvenated with regard to age-related DNAm. Overall, iPS-MSCs are similar to MSCs, but they reveal incomplete reacquisition of immunomodulatory function and MSC-specific DNAm patterns-particularly of DNAm patterns associated with tissue type and aging.
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