Regulation of Multilineage Gene Expression and Apoptosis during in vitro Expansion of Human Bone Marrow Stromal Cells with Different Cell Culture Media

Zhu, Huiyong; Miosge, Nicolai; Schulz, Jutta; Schliephake, Henning

doi:10.1159/000313417

Cited by 8 publications

(11 citation statements)

References 79 publications

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“…Thus, the high expression of Klf2 and its generally increasing expression during passaging in M1 might depend on the characteristics of M1, which not only promoted rapid proliferation but also inhibited premature differentiation of human BMSCs (Zhu et al 2010). Concurrently, Klf5 gene expression was also increased in M1; however, this increase occurred in passages 1 and 2, and decreased dramatically during subsequent passages in M1 (Fig.…”

Section: Discussionmentioning

confidence: 93%

“…It also inhibited apoptosis and senescence of hBMSCs during repeated passaging when compared with media 2 and 3 (Zhu et al 2010). Hence, we compared Klf gene expression by hBMSCs cultured in M1 with that seen in M2 and M3 by qPCR.…”

Section: Discussionmentioning

confidence: 99%

“…When the adherent cells formed colonies and grew to 80 % confluence, half of the cells harvested were used for RT-PCR, and half were subcultured until passage 4. The three media used for cultivation of hBMSCs were described previously (Zhu et al 2010). In brief, Medium 1 (M1) was composed of 58 % low-glucose (1 g/l) Dulbecco's modified Eagle's medium (DMEM; Gibco), 40 % MCDB-201 medium (Sigma-Aldrich), 19 insulin transferrin selenium (Sigma-Aldrich), 2 9 10 -8 M dexamethasone (Sigma-Aldrich), 10 -4 M ascorbic acid 2-phosphate (Sigma-Aldrich), 2 % gentamicin (Pan-Biotech GmbH, Aidenbach, Germany), 2 % fetal calf serum (FCS; Biochrom AG, Berlin, Germany), 10 ng/ml epidermal growth factor (Strathmann Biotec AG, Hamburg, Germany), 10 ng/ml platelet-derived growth factor-BB (Strathmann), 100 U penicillin, and 1,000 U streptomycin (Gibco).…”

Section: Cultivation Of Hbmscsmentioning

confidence: 99%

“…Previously, we demonstrated that culturing hBMSCs in medium M1 led to rapid proliferation without inducing differentiation, inhibited expression of apoptosis-related genes, and limited replicative senescence during repeated passaging, as compared to cells in media M2 and M3 (Zhu et al 2010). To identify Klf genes related to proliferation and pluripotency of MSCs, we compared the Klf2, Klf4, and Klf5 expression of cultures in the three media by qPCR analysis.…”

Section: Screening Of Klf2 Gene Expressionmentioning

confidence: 99%

“…In this study, using three culture media described previously (Zhu et al 2010), we determined whether Krüppel-like factors were present in cultured human bone marrow stromal cells (hBMSCs), and detected significant induction of Klf2 in hBMSCs cultured with our medium M1. We then focused on Klf2, and tested the hypothesis that it exerts a positive effect on the multipotency and proliferation of hBMSCs.…”

Section: Introductionmentioning

confidence: 99%

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Klf2 contributes to the stemness and self-renewal of human bone marrow stromal cells

Wang

Zhou

et al. 2014

Cytotechnology

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Mesenchymal stem cells (MSCs) have shown great therapeutic potential in clinical trials; however, loss of pluripotency due to culture senescence is a major factor limiting their application. Understanding the physiology of stem cell selfrenewal and stemness, and identifying the molecules that regulate these processes, are critical to future advances in tissue and organ regeneration. The Krüppel-like factor (Klf) family are key transcription factors implicated in self-renewal of embryonic stem cells. Here we identify Klf2 as a crucial transcription factor in undifferentiated human bone marrow stromal cells (hBMSCs), as indicated by gene expression in three culture media. To investigate the role of Klf2 in detail, an overexpression study using a lentiviral system in hBMSCs was performed. After Klf2 overexpression, cell proliferation was increased. The expression of pluripotency-associated genes, including Oct4, Nanog, and Rex1, was also upregulated by Klf2 overexpression. In addition, quantitative RT-PCR indicated a lower level of expression of differentiation related genes in Klf2 overexpressing cells as compared to control cells. Our results identify a functionally conserved role for Klf2 in hBMSCs, in which its expression is biologically important for stemness and self-renewal. These results are the first to show a role for Klf2 in the proliferation and pluripotency of hBMSCs.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Cultivation Of Hbmscsmentioning

confidence: 99%

Section: Screening Of Klf2 Gene Expressionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Klf2 contributes to the stemness and self-renewal of human bone marrow stromal cells

Wang

Zhou

et al. 2014

Cytotechnology

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The transfer of host MHC class I protein protects donor cells from NK cell and macrophage-mediated rejection during hematopoietic stem cell transplantation and engraftment in mice

Chow

Whiteley

et al. 2013

Stem Cells

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Human hematopoietic stem cell engraftment has been studied extensively using xenograft transplant models with immunocompromised mice. It is standard practice to incorporate mouse models, such as the limiting dilution assay, to accurately assess the number of repopulating stem cells in bone marrow or umbilical cord blood collections or to confirm the long-term repopulating ability of cultured hematopoietic stem cells. In a previous study using a standard NOD/SCID mouse model to assess human hematopoietic stem cell engraftment we observed that all human cells had mouse MHC class I protein on their surface, suggesting that this is a mechanism adopted by the cells to evade host immune surveillance. To determine whether this was a xenograft phenomenon we studied host MHC transfer in an intraspecies mouse model and observed similar results. The transfer of MHC class I proteins has implications for antigen presentation and immune modulation. In this report, we used a standard mouse model of bone marrow transplantation to demonstrate that surface protein transfer between cells plays an important role in protecting donor hematopoietic cells from NK cell and macrophage-mediated rejection. The transfer of intact MHC class I antigens from host cells to transplanted donor cells confers a self identity on these otherwise foreign cells. This gives them the ability to evade detection by the host NK cells and macrophages. Once full donor chimerism is established, transplanted cells no longer require host MHC class I protein transfer to survive.

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Epidermal growth factor can optimize a serum-free culture system for bone marrow stem cell proliferation in a miniature pig model

Wang

Zheng

Liu

et al. 2013

In Vitro Cell.Dev.Biol.-Animal

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Bone marrow-derived mesenchymal stem cells have become an attractive cell source for periodontal ligament regeneration treatment because of their potential to engraft to several tissue types after injury. Most researchers have focused on the transplantation process, but few have paid attention to cell safety concerns and rapid proliferation before transplantation. Using serum-free medium to culture stem cells may be an effective method to avoid problems associated with exogenous serum and the addition of growth factors to promote cell proliferation. Here, we randomly divided our serum-free cultures and treated them with different levels of epidermal growth factor (EGF). We then evaluated changes in rates of cell adhesion, proliferation, apoptosis, and cell cycle ratio as well as their differentiation potential. The data showed that all of these parameters were significantly different when comparing serum-free cultures with and without 10 nM/L EGF (p < 0.05/0.01); however, cells with 10 nM/L EGF did not respond differently than cells grown in standard serum-containing media without EGF (p > 0.05). In summary, our results demonstrate that 10 nM/L EGF was the optimal dose for serum-free culture, which can replace traditional standard serum medium for in vitro expansion of miniature pig bone marrow-derived mesenchymal stem cells.

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Regulation of Multilineage Gene Expression and Apoptosis during in vitro Expansion of Human Bone Marrow Stromal Cells with Different Cell Culture Media

Cited by 8 publications

References 79 publications

Klf2 contributes to the stemness and self-renewal of human bone marrow stromal cells

Klf2 contributes to the stemness and self-renewal of human bone marrow stromal cells

The transfer of host MHC class I protein protects donor cells from NK cell and macrophage-mediated rejection during hematopoietic stem cell transplantation and engraftment in mice

Epidermal growth factor can optimize a serum-free culture system for bone marrow stem cell proliferation in a miniature pig model

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