Isolation and characterization of mesenchymal stem cells from human fetus heart

Abstract

BackgroundMesenchymal stem cells (MSCs) are promising cells for cardiovascular regenerative medicine. However, their potential may be limited, because of their restricted cardiovascular differentiation potential and decline in their number and functional characteristics with increasing donor age. We have previously shown that rat fetus heart harbors primitive MSCs and administration of these cells improved left ventricular (LV) function after ischemia/reperfusion injury in rats. To evaluate their potential as a new cell type for clinical cardiovascular cell therapy, we have undertaken this study on the isolation and characterization of human fetal cardiac MSCs (hfC-MSCs).MethodsMSCs were isolated from the heart of five 14-16-week-old aborted human fetuses and studied for their growth characteristics, karyotypic stability and senescence over successive passages, expression of mesenchymal and embryonal markers by flow cytometry and immunocytochemistry, constitutive expression of cardiovascular genes by RT-PCR, differentiation into cells of the cardiovascular lineage and their immunomodulatory properties.ResultsThe hfC-MSCs grew as adherent monolayer with spindle shaped morphology and exhibited rapid proliferation with an average population doubling time of 34 hours and expansion to up to more than 80 population doublings with maintenance of a normal karyotype and without senescence. Immunophenotyping showed that they had similar phenotype as human bone marrow mesenchymal stem cells (hBM-MSCs) expressing CD73, CD90, CD105 and lacking expression of CD31, CD34, CD45, HLA-DR. However, hfC-MSCs expressed significantly higher levels of CD117 and SSEA-4 compared to hBM-MSCs. In addition, hfC-MSCs expressed the embryonal markers Oct-4, Nanog and Sox-2 as compared to hBM-MSCs. Further, hfC-MSCs had significantly higher expression of the cardiovascular genes viz. ISL-1, flk-1, GATA-4, NKX2.5 and MDR-1 as compared to hBM-MSCs, and could be differentiated into major cardiovascular cells (cardiomyocytes, endothelial cells, smooth muscle cells). Interestingly, hfC-MSCs markedly reduced T-lymphocyte proliferation with an increased secretion of TGF-β and IL-10.ConclusionsOur results show that human fetus heart is a novel source of primitive MSCs with cardiovascular commitment which may have a potential therapeutic application in cardiovascular regenerative medicine.

Single cell‐derived clonally expanded mesenchymal progenitor cells from somatic cell nuclear transfer‐derived pluripotent stem cells ameliorate the endometrial function in the uterus of a murine model with Asherman’s syndrome

“…To guarantee high purity and no teratomas of hPSC-MPCs from undifferentiated stem cells, we selected a single MPC from de- and SSEA4 were hardly detected in both hPSC-MPC-SCDs, while SSEA4 was expressed in hBM-MPCs at 46.0%, similar to that reported previously ( Figure 3D). 26 In the case of pluripotency genes, the expression levels of OCT4, SOX2 and NANOG were highly reduced during differentiation ( Figure 3E). The precursor genes for multilineage differentiated cells, precursor markers for each lineage, C/EBPα, RUNX2 and SOX9, were clearly expressed in both hPSC-MPC-SCDs.…”

Mesenchymal stem cells: Cell therapy and regeneration potential

“…(Batsali et al, 2017;Beeravolu et al, 2017;Im et al, 2005;Jin et al, 2013;Vishnubalaji et al, 2012) PB Up to P5 27 hr 50% (Jumi Kim, Shin, Jeon, Chung, & Chae, 2012) UCB Up to P18 40 hr 24% (Heo, Choi, Kim, & Kim, 2016;Jin et al, 2013;Pievani et al, 2014) CL Up to P22 54 hr 59% (Araujo et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017) WJ Up to P20 43 hr 80% (Batsali et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017;Trivanovic et al, 2013) CPJ > P40 16 hr 92% (Bakshi, McKee, Walker, Brown, & Chaudhry, 2018;Beeravolu et al, 2016;Beeravolu et al, 2017) PC Up to P10 51 hr 16% (Araujo et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017;Heo et al, 2016) FT-blood Up to P20 50 hr ND (Campagnoli et al, 2001;In't Anker et al, 2004) FT-BM Up to P20 33 hr 34% (Brady et al, 2014;Campagnoli et al, 2001;K.-S. Shin et al, 2009;Q. Wang et al, 2016) FT-heart Up to P15 80 hr ND (Garikipati et al, 2018) FT-liver Up to P20 52 hr ND (Campagnoli et al, 2001;In't Anker et al, 2004) FT-pancreas Up to P30 30 hr ND (Hu et al, 2003) AF Up to P7 39 hr 15% (Moraghebi et al, 2017) Abbreviations: AF, amniotic fluid; AT, adipose tissue; BM, bone marrow; CL, cord lining; CPJ, cord placenta junction; FT, fetal tissue; MSCs, mesenchymal stromal/stem cells; ND; not determined; P, passage; PB, peripheral blood; PC, placenta; UCB, umbilical cord blood; WJ, Wharton's jelly 2 | ADULT MSCs Bone marrow (BM) MSCs were first discovered in 1976 by Friedenstein, Gorskaja, and Kulagina (1976). BM-MSCs have been investigated...…”

Evaluation of Redox Activity of Human Myocardium‐derived Mesenchymal Stem Cells by Scanning Electrochemical Microscopy

“…The experiments were carried out in the following order: the UME was set to a negative potential and brought as close as possible to the cell's bottom surface. Afterwards, the electrochemical cell/bath was switched off immediately (voltage switching mode) . After adding a known amount of redox mediator, the media was mixed gently, a positive potential was applied and the UME was retreated at 200 μm from the cell's surface.…”