Multipotency of Flk1CD34 progenitors derived from human fetal bone marrow.
ABSTRACT We report that a cell population derived from human fetal bone marrow, termed Flk1+CD34- multipotent stem cells, can differentiate not only into osteogenic, adipogenic, and endothelial lineages but also into hepatocyte-like cells and neural and erythroid cells at the single-cell level. We depleted mononuclear cells from fetal bone marrow of CD45+, GlyA+, and CD34+ cells with the use of micromagnetic beads, then cultured them by limiting dilution. Three single colonies were harvested, expanded, and characterized. The clones have been expanded for more than 50 cell doublings, and cell-doubling time was about 30 hours. About 90% cells were in the G(0)/G(1) phase of the cell cycle, and the cells from the single colony maintained Flk1+ and CD34-. Because fetal bone marrow-derived Flk1+CD34-multipotent stem cells have the capacity for self-renewal and multilineage differentiation even after being expanded for more than 50 cell doublings, they may be an ideal source of stem cells for the treatment of inherited or degenerative diseases.
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ABSTRACT: The efficacy of stem cell transplantation for promoting recovery of patients with neurological diseases, such as stroke, has been reported in several studies. However, the safety of the intracerebral transplantation of human mesenchymal stem cells (hMSCs) remains unclear. The aim of the study was to evaluate the safety of hMSCs transplanted in cerebrum of Macaca fascicularis and to provide evidence for clinical application. A total of 24 M fascicularis were assigned to 3 groups randomly: low dose (3.0 × 10(5) cells/kg), high dose (2.5 × 10(6) cells/kg), and the control (normal saline [NS]). Human mesenchymal stem cells or NS were injected into each monkey for 2 times, with an interval of 3 weeks. The injection point was located outside of the right putamen, according to a stereotactic map and preoperative magnetic resonance imaging of the monkeys. Animal health, behavior, biophysical and biochemical parameters, and brain neurological function were routinely monitored over a 6-month period posttransplantation, and the histopathologic examinations were also performed. The results showed that local pathologic damage including local tissue necrosis and inflammation was induced after the injection. The damage of low-dose and high-dose groups was greater than that of the control group, yet over time, the damage could be repaired gradually. No major hMSCs-associated changes were induced from other indicators, and the transplantation of hMSCs in monkeys did not affect total immunoglobulin (Ig) M, total IgG, CD3, CD4, or CD8 values. We therefore conclude that transplantation of hMSCs to the cerebrum represents a safe alternative for clinical application of neurological disorders.International Journal of Toxicology 08/2014; 33(5). DOI:10.1177/1091581814545244 · 1.23 Impact Factor
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ABSTRACT: Previously, we reported that a cell population derived from human fetal bone marrow (BM), termed here Flk1(+)CD34(-) postembryonic pluripotent stem cells (PPSCs) that have the characteristics of mesenchymal stem cells (MSCs), could differentiate into ectodermal, endodermal and mesodermal cell types at the single cell level in vitro, and that these cells could also differentiate into the epithelium of liver, lung, gut, as well as the hematopoietic and endothelial lineages after transplantion into irradiated non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. In this study, we further isolated pluripotent stem cells from human fetal heart, liver, muscle, lung, derma, kidney, and fat and then analyzed the characteristics and function of these stem cells. It was found that the phenotype of the culture-expanded pluripotent stem cells from different fetal tissues was similar to BM-derived Flk1(+)CD34(-) PPSCs, i.e. Flk1 and CD44 positive, GlyA, CD34, CD45, class I-HLA and HLA-DR negative. Morphologically, these cells were fibroblast-like and the doubling time was about 30 h. More importantly, culture-expanded pluripotent stem cells from all these fetal tissues were able to differentiate into cells with morphologic and phenotypic characteristics of adipocytes, osteocytes, neurons, glial cells and hepatocytes. These pluripotent stem cells with characteristics similar to fetal BM-derived Flk1(+)CD34(-) PPSCs can be selected and cultured from tissues other than the BM. This phenomenon may help explain the "stem cell plasticity" found in multiple human tissues. In addition, as fetal BM-derived Flk1 + CD34(-) PPSCs, these pluripotent stem cells from different fetal tissues had the capacity for self-renewal and multi-lineage differentiation even after being expanded for more than 40 population doublings in vitro. Thus, they may be an ideal source of stem cells for treatment of inherited or degenerative diseases.Frontiers of Medicine in China 05/2007; 1(2):185-91. DOI:10.1007/s11684-007-0035-1
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ABSTRACT: Background: Finding factors that can increase neurogenesis are of great importance. To find these factors, it seems that neural stem cells culture is an ideal method. To analyze the effect of different factors in a limited period of time and with the least cost, finding an easier and more efficient method than normal manual counting method is needed. The aim of this study was using flow cytometry as an alternative method to evaluate neural stem cells neurogenesis. Methods: Neural stem cells from E14 mouse brain have been differentiated in a one- step and a two -step methods. After performing immunohistochemistry for neuronal and astrocytic markers, manual and flow cytometry methods have been compared in determining the percentage of neurons and astrocytes. Then, the percentage of neurons and astrocytes generated in two different differentiation methods has been compared using flow cytometry. Findings: Our findings showed that there wasn't any statistical difference between manual and flow cytometry methods in determining the percentage of neurons and astrocytes. Comparing differentiation methods by flow cytometry, showed that the percentage of both neurons and astrocytes were significantly different in theses two methods (p<0.001). Conclusion: Flow cytometry is a simple and reliable method that can replace manual counting method to evaluate neurogenesis of the neural stem cells. This method would be very useful especially when a high content screening of different factors and compounds is needed. Key words: Neural stem cell, differentiation, flow cytometry, neurogenesis