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

(6 citation statements)

References 27 publications

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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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“…Compared to primary MSCs, cultured MSCs with high levels of CD44 displayed decreased targeting to the bone marrow [13]. Our previous research revealed that stem cell antigen 1 (Sca-1) is expressed at higher levels in adherent cultured mouse MSCs (AC-mMSCs) compared to mMSCs in nonadherent cultures maintained in ultra-low-adherent plates (nonAC-mMSCs) [14]. Sca-1 + mMSCs play a crucial role in improving cardiac function in MI [14,15].…”

Section: Introductionmentioning

confidence: 99%

“…Our previous research revealed that stem cell antigen 1 (Sca-1) is expressed at higher levels in adherent cultured mouse MSCs (AC-mMSCs) compared to mMSCs in nonadherent cultures maintained in ultra-low-adherent plates (nonAC-mMSCs) [14]. Sca-1 + mMSCs play a crucial role in improving cardiac function in MI [14,15]. Compared with nonadherent cultured human MSCs (nonAC-hMSCs), adherent cultured hMSCs (AC-hMSCs) with high protein expression of caspase-3, caspase-7, and caspase-9 had a marked decrease in cell apoptosis [16].…”

Section: Introductionmentioning

confidence: 99%

Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway

et al. 2020

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Background Mesenchymal stem cells (MSCs) can provide therapeutic benefits for myocardial infarction (MI) recovery; however, the molecular mechanism by which MSCs improve the heart function is unclear. Methods Microarray analysis was performed to examine the expression profiling of human MSCs (hMSCs) grown as adherent cultures (AC-hMSCs) or nonadherent cultures on ultra-low-adherent plates (nonAC-hMSCs). Real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, and enzyme-linked immunosorbent assays (ELISA) were used to assess VEGFA expression and secretion in the AC-hMSCs and nonAC-hMSCs. The paracrine effect of VEGFA-overexpressing AC-MSCs (AC-VEGFA-hMSCs) or VEGFA-knockdown nonAC-hMSCs (nonAC-shVEGFA-hMSCs) on the angiogenic ability of human umbilical vein endothelial cells (HUVECs) was evaluated using tube formation assay. AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs were transplanted into myocardial infarction rats to investigate the therapeutic effect of AC-VEGFA-hMSCs or nonAC-shVEGFA-hMSCs. Luciferase reporter assay was used to confirm the association of VEGFA with miR-519d. Results Microarray analysis revealed that VEGFA is downregulated in AC-hMSCs compared to nonAC-hMSCs. Functional assays revealed that high levels of VEGFA produced from AC-VEGFA-hMSCs increased the tube formation capacity of HUVECs in vitro, improved angiogenesis and cardiac performance, and reduced infarct size in a rat MI model. Low levels of VEGFA secretion from nonAC-shVEGFA-hMSCs had the opposite effects. Mechanistically, we found that miR-519d directly targets VEGFA. High levels of VEGFA secreted from VEGFA-overexpressing nonAC-hMSCs abolished the repressive effect of miR-519d on HUVEC angiogenesis. Conclusion Our findings indicate that nonadherent culture-induced secretion of VEGFA plays an important role in MSCs via the miR-519d/VEGFA pathway and may provide a novel therapeutic strategy for MI treatment.

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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

Nonadherent culture method promotes MSC-mediated vascularization in myocardial infarction via miR-519d/VEGFA pathway

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