Article

Mesenchymal progenitor cells in the human umbilical cord

Sungkyunkwan University, Sŏul, Seoul, South Korea
Annals of Hematology (Impact Factor: 2.4). 01/2005; 83(12):733-8. DOI: 10.1007/s00277-004-0918-z
Source: PubMed

ABSTRACT Mesenchymal progenitor or stem cells (MPCs) isolated from fetal blood, liver, and bone marrow are a population of multipotential cells that can proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat, and stroma. The objective of this study was to isolate and characterize MPCs in the human umbilical cord. The suspensions of endothelial and subendothelial cells in cord vein were collected and cultured in M199 supplemented with 10% fetal bovine serum (FBS). Of 50 umbilical cord samples, 3 had numerous fibroblastoid cells morphologically distinguishable from endothelial cells. Fibroblastic cells displayed lack of expression of vWF, Flk-1, and PECAM-1, indicating the endothelial cell-specific marker. To investigate the differentiation potentials, the cells were cultured in adipogenic or osteogenic medium for 2 weeks. Fibroblast-like cells treated with adipogenic supplementation showed Oil red O-positive staining and expressed adipsin, FABP4, LPL, and PPARgamma2 genes by reverse transcriptase polymerase chain reaction (RT-PCR). In osteogenic differentiation, alkaline phosphatase activity and calcium accumulation were detected. RT-PCR studies determined that Cx43, osteopontin, and Runx2 genes were expressed in the osteogenic cultures. Among three cell lines cultured continuously for passage 10, two had normal karyotypes; however, one retained a karyotype of mos 46,XY[19]/47,XY,+mar[3]. These observations suggest that MPCs are present in human umbilical cord and possess several typical traits of MPCs.

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    • "First was a standard keratocyte medium (KM), [17,46] consisting of DMEM (Gibco, Invitrogen, Paisley, UK), supplemented with 10% vol/vol heat-inactivated FBS (Fisher Scientific), 0.02 μg/ml gentamicin, 0.5 ng/ml amphotericin B (Gibco), 4.5 μg/ml insulin, human recombinant (Gibco), and 0.5% vol/vol DMSO (Sigma Aldrich). The second was a medium previously shown to support the expansion of MSCs [15,61-63] (MM), consisting of M199 medium (Sigma) supplemented with 20% vol/vol heat-inactivated FBS, 2.5 μg/ml antibiotic solution, Plasmocin (Autogen Bioclear, Wiltshire, UK), 0.02 μg/ml gentamicin, 0.5 ng/ml amphotericin B (Gibco), and 1.59 mM L-glutamine (Sigma Aldrich). "
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    ABSTRACT: The corneal stroma is being increasingly recognized as a repository for stem cells. Like the limbal and endothelial niches, stromal stem cells often reside in the peripheral cornea and limbus. These peripheral and limbal corneal stromal cells (PLCSC) are known to produce mesenchymal stem cells in vitro. Recently, a common corneal stromal and epithelial progenitor has been hinted at. This study aims to examine the stem cell potential of corneal stromal cells and to investigate their epithelial transdifferentiation ability. PLCSC were grown in traditional Dulbecco's modified Eagle's medium (DMEM)-based keratocyte culture medium and an M199-based medium and analysed for a profile of cell surface markers using flow cytometry and differentiated into mesenchymal phenotypes analysed by qPCR and histological staining. PLCSC in M199 were subsequently divided into subpopulations based on CD34 and CD105 expression using fluorescent activated cell sorting (FACS). Subpopulations were characterized by marker profile and mesenchymal differentiation ability. Both whole PLCSC and subpopulations were also cultured for epithelial transdifferentiation. Cells cultured in M199 demonstrated a more stem-like cell surface marker profile and the keratocyte marker CD34 was retained for several passages but absent in cells cultured in DMEM. Cells cultured in M199 also exhibited a greater mesenchymal differentiation potential, compared with DMEM. PLCSC could be divided into CD34+CD105+, CD34-CD105+ and CD34-CD105- subpopulations, of which CD34+CD105+ cells were the most stem-like with regard to marker expression and mesenchymal differentiation potential. Subpopulations of PLCSC exhibited differing abilities to transdifferentiate into epithelial phenotypes. Cells that were initially CD34+CD105+ showed greatest differentiation potential producing CK3+ and CK19+ cells, and expressed a range of both epithelial progenitor (HES1, FRZB1, DCT, SOD2, ABCG2, CDH1, KRT19) and terminally differentiated (DSG3, KRT3, KRT12, KRT24) genes. Culture medium has a significant effect on the phenotype and differentiation capacity of PLCSC. The stroma contains a heterogeneous cell population in which we have identified CD34+ cells as a stem cell population with a capacity for mesenchymal and epithelial differentiation.
    Stem Cell Research & Therapy 06/2013; 4(3):75. DOI:10.1186/scrt226 · 4.63 Impact Factor
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    • "UCMSCs isolated from cord tissue samples processed after 04 days and 30 days of collection were incubated with growth medium containing 0.25mg of colcemid. After 4 hours of incubation the cells were harvested and resuspended in 0.075M KCl and then fixed in 3:1 methanol/acetic acid.11 GTG banding was done on metaphase spreads obtained from cultured UCMSCs. "
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    • "Human umbilical cord blood cells (HUCBCs) are a source of hematopoietic stem cells, endothelial cell precursors, mesenchymal progenitors, and other multipotent/ pluripotent lineage stem cells and represent a promising option for alternative for experimental stroke therapies (Chen et al., 2001, 2006; Kim et al., 2004; Vendrame et al., 2004, 2005; Boltze et al., 2005, 2011b; Newman et al., 2005; Berger et al., 2006; Newcomb et al., 2006; Bewley and Mercer, 2010). One of the contributing factors for their therapeutic efficacy for experimental stroke is that HUCBCs provide a ready supply of neurotrophic and angiogenic factors, and induce neurogenesis and angiogenesis (Chen et al., 2001, 2007; Hau et al., 2008; Jiang et al., 2008; Chung et al., 2009; Liu et al., 2009; Park et al., 2009; Arien-Zakay et al., 2011; Terry et al., 2011; Nih et al., 2012). "
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    Neuroscience 10/2012; 227. DOI:10.1016/j.neuroscience.2012.09.066 · 3.33 Impact Factor
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