| Background: Mesenchymal stem cells (MSCs) are multipotent stem cells ofself-renewal and multidifferentiation potential. Although MSCs can be isolated fromseveral tissues such as bone marrow, umbilical cord, adipose tissue, cord blood, andsynovial membrane, MSCs are the richest in the umbilical cord.Objective: To investigate the identification of human umblical cord derivedmesenchymal stem cells isolated and expanded.Methods:(1) hUC-MSCs were isolated by two enzymes digestion;(2)Immunophenotypes of hUC-MSCs were analyzed by flow cytometry;(3)Adipogenic and osteogenic differentiation of hUC-MSCs were observed by oil red Ostaining and Alizarin red S staining.Results:(1) hUC-MSCs were typically fibroblast-like, plastic-adherent cells;(2)hUC-MSCs expressed high levels of mesenchymal immunophenotypes such asCD29, CD44, CD54, CD73, CD90, CD105,CD106and HLA-ABC, did not expresshematopoietic immunophenotypes such as CD11b, CD14, CD34, CD45, HLA-DRand costimulatory molecules CD80, CD86;(3) hUC-MSCs had adipogenic andosteogenic differentiation potential.Conclusion: We had isolated and expanded successfully human umblical cordderived mesenchymal stem cells. Background: MSCs have powerful immunomodulation properties, they canregulate the functions of several immune cells such as T cells, B cells, natural killercells, dendritic cells, antigen presenting cells, neutrophils and macrophages.Serum-free media support the expansion of MSCs, maintaining themselves sizes,immunophenotypes and differentiation of cells in vitro. However, the effect ofserum-free media on immunoregulatory properties of UC-MSCs in vitro is notunclear.Objective: To investigate the effect of serum-free media on immunoregulatoryproperties of hUC-MSCs in vitro.Methods:(1)Immunophenotypes of hUC-MSCs and cell apoptosis wereanalyzed by flow cytometry;(2) Adipogenic and osteogenic differentiation ofhUC-MSCs were observed by oil red O and Alizarin red S staining;(3) MTSmeasured cells proliferation;(4) IFN-γ and PGE2levels in cell free supernatants werederminated by ELISA; the mRNA levels of IFN-γ, TNF-α, IL-21, IL-22,IL-26, IDO1,COX2, HGF, IL-1β and IL-6were tested by real time-PCR.Results:(1) hUC-MSCs expanded in serum-free media were fibroblast-like,plastic-adherent cells; expressed high levels of mesenchymal surface markers CD29,CD44, CD73, CD90, CD105, CD106, and low levels of hematopoietic surfacemarkers CD34; hUC-MSCs in serum-free media had adipogenic and osteogenicdifferentiation;(2) hUC-MSCs in serum-free media inhibited the proliferation andapoptosis of CD4+T cells;(3) hUC-MSCs in serum-free media supressed CD4+Tcells secreting IFN-γ and expressing the mRNA of IFN-γ, TNF-α, IL-21, IL-22, andIL-26;(4) hUC-MSCs in serum-free media secreted high levels of PGE2, andexpressed high levels of IDO1, COX2, IL-1β, and IL-6mRNA, didn’t express HGF.Conclusion:(1) hUC-MSCs cultured in serum-free media maintained themselves biological characteristics of morphologes, adipogenic and osteogenicdifferentiation, and immunophenotypes.(2) Compared with hUC-MSCs in serum-containing media, hUC-MSCs inserum-free media could also inhibit the proliferation and apoptosis of CD4+T cells;(3) hUC-MSCs in serum-free media also had powerful immunosuppressiveactivities and secreted high levels of immunomodulatory factors as well as those inserum-containing media. Background: Pulmonary arterial hypertension (PAH) is a progressive diseasecharacterized by the structural alterations of pulmonary vascular causing highblood pressure of the pulmonary circulation and leading to right heart failure. PAHis a common disease, the pathogenesis of it is not clear. But studies found that theimmnue factors play an critical role in the pathogenesis of PAH.Objective: In order to further evaluate the immunomodulation properties ofhUC-MSCs in serum free medium in vivo.Methods:(1) The pulmonary arterial hypertension model was induced bymonocrotaline：ethanol saline dissolved monocrotaline, the dose of50mg/kg wasinjected into back subcutaneous tissue of rat;(2) Mean pulmonary arterial pressurewas tested by polystyrene catheter;(3) Structural alterations of pulmonary tissueswere determined by hematoxylin and eosin (H&E) staining;(4) hUC-MSCs werelabeled by CM-DiI;(5) The TNF-α levels of peripherial blood and lung tissues weretested by ELISA; The mRNA expressions of TNF-α, IFN-γ, IL-1β, IL-6and IL-18inlung tissues were determinated by real time PCR.Results:(1) The hair, respiration, and activities were not abnormal obviously inrats transplated with hUC-MSCs in serum-free media and the weights of rats wereincreased;(2) hUC-MSCs in serum-free media decreased significantly the meanpulmonary arterial pressure;(3) hUC-MSCs in serum-free media attenuatedsignificantly pulmonary vascular remodeling;(4)There were lots of CM-DiI labeledmesenchymal stem cells in the lung tissues of rats transplanted with hUC-MSCs inserum-free media.(5) hUC-MSCs in serum-free media decreased obviously theTNF-α protein levels of peripherial blood and lung tissues and the mRNA expressions of TNF-α, IFN-γ, IL-1β, IL-6and IL-18of lung tissues.Conclusion:(1) Compared with hUC-MSCs in serum-containing media,hUC-MSCs in serum-free media could also attenuate significantly pulmonaryvascular remodeling.(2) hUC-MSCs in serum-free media also had powerful immunosuppressiveactivities as well as those in serum-containing media in vivo;... |
