Nonadherent culture method downregulates stem cell antigen-1 expression in mouse bone marrow mesenchymal stem cells

Deng, Baoping; Deng, Weiping; Xiao, Pengnan; Zeng, Kuan; Zhang, Shining; Zhang, Hongwu; Deng, David Y.B.; Yang, Yanqi

doi:10.3892/etm.2015.2457

Cited by 7 publications

(3 citation statements)

References 27 publications

(31 reference statements)

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“…It is also possible that HSC may be present, but after for 3–4 weeks of culture, attain a different phenotype. In particular, culturing of Lin - Sca-1 + c-Kit + cells over stroma may lead to loss of the Sca-1 marker, a phenomenon reported previously for mesenchymal stem cells in vitro (Deng et al, 2015). Previously we reported evidence for maintenance of low numbers of HSPC in spleen longterm stromal cultures through ability to induce hematopoietic reconstitution in irradiated host mice (O’Neill et al, 2014).…”

Section: Resultsmentioning

confidence: 58%

Identification of Stromal Cells in Spleen Which Support Myelopoiesis

Lim

O’Neill

2019

Front. Cell Dev. Biol.

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Stromal cells in spleen organize tissue into red pulp, white pulp and marginal zone, and also interact with hematopoietic cells to regulate immune responses. This study has used phenotypic information of a previously described spleen stromal cell line called 5G3, which supports restricted hematopoiesis in vitro, to identify an equivalent stromal cell subset in vivo and to test its capacity to support hematopoiesis. Using stromal cell fractionation, phenotypic analysis, as well as cell growth and hematopoietic support assays, the Sca-1+gp38+Thy1.2+CD29+CD51+ fraction of spleen stroma has been identified as an equivalent stromal subset resembling the 5G3 cell counterpart. While heterogeneity may still exist within that subset, it has been shown to have superior hematopoietic support capacity compared with the 5G3 cell line, and all other spleen stromal cell fractions tested.

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Section: Resultsmentioning

confidence: 58%

Identification of Stromal Cells in Spleen Which Support Myelopoiesis

Lim

O’Neill

2019

Front. Cell Dev. Biol.

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“…The major reason for lacking information about Ly6a in our scRNA-seq data is that we cultured the periosteal cells for a week to acquire enough cells for scRNA-seq. It has been reported that adherent cell culture could downregulate the expression of Ly6a in mesenchymal stem cells (Deng et al 2015). Thus, we chose to label resident Ctsk + Ly6a + cells in vivo with immunofluorescent staining to exhibit their specific distribution in the periosteum of jawbone.…”

Section: Discussionmentioning

confidence: 99%

Identification of Periosteal Osteogenic Progenitors in Jawbone

Ding

Geng

et al. 2022

J Dent Res

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Unlike long bones, jawbone development is mainly accomplished by intramembranous ossification resulting from the differentiation of periosteal progenitor cells. However, the spatiotemporal ontogeny of periosteal progenitor cells during jawbone development and repair remains elusive. In this study, we mapped the transcriptional landscape of the human jawbone periosteum at single-cell resolution and identified a cathepsin K (Ctsk)+ periosteal subset. Lineage tracing analysis indicated that Ctsk-Cre–labeled periosteal cells could make contributions to jawbone development. However, different from the periosteal-specific location of Ctsk+ cells in long bone, we also identified Ctsk+ stromal cells in jawbone marrow and implied the heterogeneity of jawbone Ctsk+ hierarchy. In further analysis of the periosteal progenitor cell subset of heterogeneous Ctsk+ hierarchy, we identified a unique Ctsk+Ly6a+ subset of cells. The additional marker Ly6a helped to further confine the progenitor subset to the jawbone periosteum and was nearly undetectable in the bone marrow. Defects in the jawbone could activate the migration and osteogenic differentiation of Ctsk+Ly6a+ cells. Local ablation of Ctsk+ cells by diphtheria reduced the number of Ctsk+Ly6a+ cells and delayed the repair of the bone defect. Taken together, we identify a novel periosteal osteogenic progenitor subset that is active in jawbone osteogenesis and healing.

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“…The plastic adherent condition is considered a suitable niche for amplification of MSCs in vitro , promoting cells survival. To determine the effect of culture conditions on apoptosis of human MSCs (hMSCs), hMSCs were cultured in vitro either in ultra-low-adherence culture plates (to mimic nonadherent conditions) or in standard tissue culture plates (to mimic adherent conditions), as previously described ( 9 ). Apoptosis was then analyzed by flow cytometry.…”

Section: Introductionmentioning

confidence: 99%

Removal from adherent culture contributes to apoptosis in human bone marrow mesenchymal stem cells

Deng

Jiang

Zeng

et al. 2017

Molecular Medicine Reports

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Mesenchymal stem cells (MSCs) are routinely isolated due to their adherence to tissue culture plates and their in vitro growth characteristics. Expansion of MSCs in adherent cultures is the only way to obtain sufficient cells for use in either clinical or research settings. MSCs have tremendous potential in myocardial repair treatment by cell therapy techniques, however, a large number of MSCs die from apoptosis following transplantation. Previous studies have examined the factors contributing to the survival of transplanted cells, but little is known about the effect of removal from adherent culture conditions on apoptosis of the MSCs. In the present study, human bone marrow MSCs were expanded in adherent cultures. Then apoptosis rates were examined at different time points in MSCs cultured in nonadherent conditions (ultra-low-adherence plates) compared with MSCs cultured in adherent conditions (standard tissue culture plates). Flow cytometry analysis suggested that cell apoptosis increased when MSCs were cultured in nonadherent culture conditions. In addition, western blot and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that caspase-3, −7 and −9 were involved in this process. The present study demonstrated that loss of culture adherence increases apoptosis of human MSCs. The present findings may provide new insight into the factors affecting MSC survival after transplantation.

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Nonadherent culture method downregulates stem cell antigen-1 expression in mouse bone marrow mesenchymal stem cells

Cited by 7 publications

References 27 publications

Identification of Stromal Cells in Spleen Which Support Myelopoiesis

Identification of Stromal Cells in Spleen Which Support Myelopoiesis

Identification of Periosteal Osteogenic Progenitors in Jawbone

Removal from adherent culture contributes to apoptosis in human bone marrow mesenchymal stem cells

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