scholarly journals Long-term expansion enhances the expression of tumor suppressor genes in human bone marrow-derived mesenchymal stem cells

2019 ◽  
Vol 22 (1) ◽  
pp. 136-142
Author(s):  
Loan Thi Tung Dang ◽  
Anh Thi Van Bui ◽  
Nhat Chau Truong ◽  
Huy Duc Van ◽  
Phuc Van Pham

Introduction: Mesenchymal stem cells (MSCs) are possibly the most potent type of stem cells for the treatment of many diseases since they possess many advantageous properties, such as abundant source, ease of isolation, and potential to differentiate and trans-differentiate into different types of cells. Although the therapeutic potential of expanded MSCs has been well proven, their biosafety features have not been fully understood. This study aimed to investigate some changes in phenotype and gene expression of bone marrow derived MSCs after long term expansion. Methods: In this study, expanded mesenchymal stem cells derived from human bone marrow (hBMSCs) were identified for their characteristics (which included morphology, immunophenotype, and differentiation potential) at passages 5, 10 and 15. Moreover, they were evaluated for the expression of various tumor suppressor genes (PTEN, p16, and p53) by real-time RT-PCR. Results: The results showed that the hBMSCs at passage 15 displayed a change in morphology and a slight reduction of the expression of CD44 and CD90, whereas their potential for adipogenic and osteogenic differentiation was maintained. Moreover, the expression of tumor suppressor genes in the hBMSCs increased after long-term culture. Conclusion: It could be assumed that prolonged cultures of more than 15 passages drove the hBMSCs into senescence phase. Cultured hBMSCs below passage 10 seemed to be more effective in application because their properties were still preserved.  

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Angela Bentivegna ◽  
Gaia Roversi ◽  
Gabriele Riva ◽  
Laura Paoletta ◽  
Serena Redaelli ◽  
...  

Human bone marrow mesenchymal stem cells (hBM-MSCs) are the best characterized multipotent adult stem cells. Their self-renewal capacity, multilineage differentiation potential, and immunomodulatory properties have indicated that they can be used in many clinical therapies. In a previous work we studied the DNA methylation levels of hBM-MSC genomic DNA in order to delineate a kind of methylation signature specific for early and late passages of culture. In the present work we focused on the modification of the methylation profiles of the X chromosome and imprinted loci, as sites expected to be more stable than whole genome. We propose a model where cultured hBM-MSCs undergo random modifications at the methylation level of most CGIs, nevertheless reflecting the original methylation status. We also pointed out global genome-wide demethylation connected to the long-term culture and senescence. Modification at CGIs promoters of specific genes could be related to the decrease in adipogenic differentiation potential. In conclusion, we showed important changes in CGIs methylation due to long-termin vitroculture that may affect the differentiation potential of hBM-MSCs. Therefore it is necessary to optimize the experimental conditions forin vitroexpansion in order to minimize these epigenetic changes and to standardize safer procedures.


2021 ◽  
Vol 41 ◽  
pp. 603-615
Author(s):  
A-N Zeller ◽  
◽  
M Selle ◽  
Z Gong ◽  
M Winkelmann ◽  
...  

Underlying pathomechanisms of osteoporosis are still not fully elucidated. Cell-based therapy approaches pose new possibilities to treat osteoporosis and its complications. The aim of this study was to quantify differences in human bone marrow-derived mesenchymal stem cells (hBMSCs) between healthy donors and those suffering from clinically manifest osteoporosis. Cell samples of seven donors for each group were selected retrospectively from the hBMSC cell bank of the Trauma Department of Hannover Medical School. Cells were evaluated for their adipogenic, osteogenic and chondrogenic differentiation potential, for their proliferation potential and expression of surface antigens. Furthermore, a RT2 Osteoporosis Profiler PCR array, as well as quantitative real-time PCR were carried out to evaluate changes in gene expression. Cultivated hBMSCs from osteoporotic donors showed significantly lower cell surface expression of CD274 (4.98 % ± 2.38 %) than those from the control group (26.03 % ± 13.39 %; p = 0.007), as assessed by flow cytometry. In osteoporotic patients, genes involved in inhibition of the anabolic WNT signalling pathway and those associated with stimulation of bone resorption were significantly upregulated. Apart from these changes, no significant differences were found for the other cell surface antigens, adipogenic, osteogenic and chondrogenic differentiation ability as well as proliferation potential. These findings supported the theory of an influence of CD274 on the regulation of bone metabolism. CD274 might be a promising target for further investigations of the pathogenesis of osteoporosis and of cell-based therapies involving MSCs.


2017 ◽  
Vol 118 (10) ◽  
pp. 3072-3079 ◽  
Author(s):  
Annelise Pezzi ◽  
Bruna Amorin ◽  
Álvaro Laureano ◽  
Vanessa Valim ◽  
Alice Dahmer ◽  
...  

2014 ◽  
Vol 55 (4) ◽  
pp. 845-853 ◽  
Author(s):  
Faezeh Faghihi ◽  
Esmaeil Mirzaei ◽  
Arash Sarveazad ◽  
Jafar Ai ◽  
Somayeh Ebrahimi Barough ◽  
...  

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