Efficient Derivation and Concise Gene Expression Profiling of Human Embryonic Stem Cell-Derived Mesenchymal Progenitors (EMPs)

Abstract

New potential sources of stem cells for clinical application include bone marrow mesenchymal stem cells (BMMSCs), human embryonic stem cells (hESCs), and induced pluripotent stem cells (iPS). However, each source is not without its own concerns. While research continues in an effort to overcome these problems, the generation of mesenchymal progenitors from existing hESC lines may circumvent many of these issues. We report here a simple and efficient method of generating hESC-derived mesenchymal progenitors (EMPs) and transcriptome profiling using a concise, custom-designed, oligomnucleotide gene expression microarray. Characterization of EMPs shows that these cells are similar to BMMSCs in terms of differentiation capacity as well as cell surface marker expression. In addition, EMPs express several ESC markers and HLA-G, a nonclassical MHC class I molecule with immunomodulatory properties. Morevoer, EMPs possess significantly enhanced proliferative ability over BMMSCs during which karyotypic stability was maintained. Although derived from hESCs, EMPs do not form any tumors in immunocompromised mice. To efficiently profile gene expression in multiple samples, we designed an oligoarray to probe just over 11,000 genes highly expressed in stem cells. We found that the transcriptome of EMPs is more similar to BMMSCs than hESCs. Both cell types highly express genes involved in processes related to the cytoskeleton, extracellular matrix, and cell adhesion, but EMPs show higher expression of genes involved in cell proliferation whereas BMMSCs showed higher expression of immune-related genes. Based on our data, EMPs may be an accessible source of mesenchymal progenitor for therapeutic use.

Functional Comparison of Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Cells and Bone Marrow-Derived Mesenchymal Stromal Cells from the Same Donor

“…[10,39] on the basis of experiments showing that these PSC derivatives satisfied the minimal MSC criteria put forth by the ISCT [22,23]. On the other hand, recent published studies based on gene expression data [10,15,40] and in vitro differentiation assays [29,39,[41][42][43] 1604 DIEDERICHS AND TUAN suggested differences between BMSCs and MSC-like PSC derivatives. However, when interpreting these studies, it should be taken into consideration that the MSCs and ESCs originated from different donors.…”

Differentiation behavior of iPS cells cultured on PLGA with osteoinduction medium

“…Repeated passaging by trypsinization has been showed to enriched MSCs population 28) . But, mechanism of why functional MSCs, or mesenchymal progenitor cells named by Yen et al in their study, can efficiently derived from iPSCs by serial culturing and trypsinization needs further investigations 29) . Zou et al showed one possibility, it might be associated with the used serum, but speculated that the serum effect should not be main effector 26) .…”

Age-Related Decreases of Serum-Response Factor Levels in Human Mesenchymal Stem Cells Are Involved in Skeletal Muscle Differentiation and Engraftment Capacity

“…S1; Supplementary Data are available online at www.liebertpub.com/scd) [9,11,21,22,29,30]. The ability of hDE-MSCs to differentiate into various myogenic lineages, however, has not been investigated.…”

“…hDE-MSCs, as a type of MSCs, are capable of multilineage differentiation [9,11,21,22,29,30], and we sought to ascertain the specificity of MM-induced SkM differentiation in these MSCs by assaying for changes in the levels of Runx2 and PPAR-c, the master lineage transcription factors for osteogenesis and adipogenesis, respectively [40,41]. We found that in all types of hDEMSCs undergoing SkM differentiation, the expression levels of Runx2 and PPAR-c are decreased compared with undifferentiated hDE-MSCs (Supplementary Fig.…”

“…MSCs were derived from HSF-6 human ESCs (NIH code UC06, obtained from the University of California, San Francisco under a Materials Transfer Agreement) as previously reported [20,21]. Placenta-derived MSCs were isolated from term placenta obtained from healthy mothers with informed consent obtained as approved by the institutional review board [11].…”