Telomerase reverse transcriptase mediated immortalization of human bone marrow stromal cells

Teng, Yue; Hu, Yunyu; Li, Xu Sheng; Wang, Zhen; Wang, Rui

doi:10.1590/s1516-89132014000100006

Cited by 2 publications

(2 citation statements)

References 15 publications

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“…However, despite this, we noted a variable response with respect to proliferative potential where 1C6H was the only cell type to undergo uninterrupted proliferation. The minimal changes in proliferative capacity seen in BMA13H are in contrast to most other publications reporting successful immortalisation of MSCs[ 54 – 56 ] although some report similar results to ours with a small gain in proliferation capacity but not immortalisation[ 57 ]. The previous track record for immortalisation success indicates that the cessation of growth in our cells may be due to a mechanism other than telomere shortening.…”

Section: Discussionsupporting

confidence: 76%

Immortalisation with hTERT Impacts on Sulphated Glycosaminoglycan Secretion and Immunophenotype in a Variable and Cell Specific Manner

et al. 2015

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BackgroundLimited options for the treatment of cartilage damage have driven the development of tissue engineered or cell therapy alternatives reliant on ex vivo cell expansion. The study of chondrogenesis in primary cells is difficult due to progressive cellular aging and senescence. Immortalisation via the reintroduction of the catalytic component of telomerase, hTERT, could allow repeated, longitudinal studies to be performed while bypassing senescent phenotypes.MethodsThree human cell types: bone marrow-derived stromal cells (BMA13), embryonic stem cell-derived (1C6) and chondrocytes (OK3) were transduced with hTERT (BMA13H, 1C6H and OK3H) and proliferation, surface marker expression and tri-lineage differentiation capacity determined. The sulphated glycosaminoglycan (sGAG) content of the monolayer and spent media was quantified in maintenance media (MM) and pro-chondrogenic media (PChM) and normalised to DNA.Results hTERT expression was confirmed in transduced cells with proliferation enhancement in 1C6H and OK3H cells but not BMA13H. All cells were negative for leukocyte markers (CD19, CD34, CD45) and CD73 positive. CD14 was expressed at low levels on OK3 and OK3H and HLA-DR on BMA13 (84.8%). CD90 was high for BMA13 (84.9%) and OK3 (97.3%) and moderate for 1C6 (56.7%), expression was reduced in BMA13H (33.7%) and 1C6H (1.6%). CD105 levels varied (BMA13 87.7%, 1C6 8.2%, OK3 43.3%) and underwent reduction in OK3H (25.1%). 1C6 and BMA13 demonstrated osteogenic and adipogenic differentiation but mineralised matrix and lipid accumulation appeared reduced post hTERT transduction. Chondrogenic differentiation resulted in increased monolayer-associated sGAG in all primary cells and 1C6H (p<0.001), and BMA13H (p<0.05). In contrast OK3H demonstrated reduced monolayer-associated sGAG in PChM (p<0.001). Media-associated sGAG accounted for ≥55% (PChM-1C6) and ≥74% (MM-1C6H).ConclusionIn conclusion, hTERT transduction could, but did not always, prevent senescence and cell phenotype, including differentiation potential, was affected in a variable manner. As such, these cells are not a direct substitute for primary cells in cartilage regeneration research.

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Section: Discussionsupporting

confidence: 76%

Immortalisation with hTERT Impacts on Sulphated Glycosaminoglycan Secretion and Immunophenotype in a Variable and Cell Specific Manner

et al. 2015

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“…Therefore, telomere length is important in guaranteeing cell proliferation stability. Enhancing telomerase activity by introducing the exogenous TERT gene into targeted cells is the primary method used in recent cell immortalization studies (Kaloyianni et al, 2015, Teng et al, 2014, Tsai et al, 2010, Wongkajornsilp et al, 2012). Hamada et al, constructed an hMSC-TERT cell line in 2003, which had biological characteristics that were no different than the original generation of hMSCs, but the detailed molecular mechanism and the function of telomerase remain unclear.…”

Section: Discussionmentioning

confidence: 99%

Derivation and characterization of sheep bone marrow-derived mesenchymal stem cells induced with telomerase reverse transcriptase

Zhu

Liu

Deng

et al. 2017

Saudi Journal of Biological Sciences

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Bone marrow mesenchymal stem cells (BMSCs) are a type of adult stem cells with a wide range of potential applications. However, BMSCs have a limited life cycle under normal culturing conditions, which has hindered further study and application. Many studies have confirmed that cells modified by telomerase reverse transcriptase (TERT) can maintain the ability to proliferate over a long period of time. In this study, we constructed a gene expression vector to transfer TERT into sheep BMSCs, and evaluated whether the TERT cell strain was successfully transferred. The abilities of cell proliferation and differentiation were evaluated using the methods including growth curve determination, inheritance stability analysis, multi-directional induction and so on, and the results showed that the cell strain can be cultured to 40 generations, with a normal karyotype rate maintained at 88.24%, and that the cell strain can be transferred and differentiated into neurocytes and lipocytes, proving that it retains the multi-directional transdifferentiation ability.

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Telomerase reverse transcriptase mediated immortalization of human bone marrow stromal cells

Abstract: Primary human bone marrow stromal cells (hMSCs) were transfected with human telomerase reverse transcriptase

Cited by 2 publications

References 15 publications

Immortalisation with hTERT Impacts on Sulphated Glycosaminoglycan Secretion and Immunophenotype in a Variable and Cell Specific Manner

Immortalisation with hTERT Impacts on Sulphated Glycosaminoglycan Secretion and Immunophenotype in a Variable and Cell Specific Manner

Derivation and characterization of sheep bone marrow-derived mesenchymal stem cells induced with telomerase reverse transcriptase

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