Hypoxia depletes contaminating CD45+ hematopoietic cells from murine bone marrow stromal cell (BMSC) cultures: Methods for BMSC culture purification

Guo, Wendi; Spiller, Kassandra; Tang, Junyao; Karner, Courtney M.; Hilton, Matthew J.; Wu, Chia‐Lung

doi:10.1016/j.scr.2021.102317

Cited by 5 publications

(4 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The Colony Forming Unit – Fibroblastic (CFU-F) assay was performed as published, with the exception of placing cells in hypoxia based on a recent publication demonstrating reduction of contaminating cells ( Guo et al, 2020 ; Robey et al, 2014 ). In short, cells were isolated from bisected tibia and femur of 2-month-old Col10a1Cre; R26-tdTomato mice centrifuged at 4°C twice pulsed up to 5 K xG.…”

Section: Methodsmentioning

confidence: 99%

“…The Colony Forming Unit -Fibroblastic (CFU-F) assay was performed as published, with the exception of placing cells in hypoxia based on a recent publication demonstrating reduction of contaminating cells. (Guo et al, 2021;Robey et al, 2014) In short, cells were isolated from bisected tibia and femur of 2M old Col10a1Cre; R26-tdTomato mice centrifuged at 4C twice pulsed up to 5K xG. Following red blood cell lysis (Roche), cells were plated at 750K cells per 15cm tissue culture plate in complete media (a-MEM, 20% FBS, 1% penicillinstreptomycin) and glutamax (Gibco) after straining through a 70um filter.…”

Section: Colony Forming Unit Assaysmentioning

confidence: 99%

See 1 more Smart Citation

Hypertrophic chondrocytes serve as a reservoir for marrow-associated skeletal stem and progenitor cells, osteoblasts, and adipocytes during skeletal development

Long

Leinroth

Liao

et al. 2022

eLife

Self Cite

View full text Add to dashboard Cite

Hypertrophic chondrocytes give rise to osteoblasts during skeletal development; however, the process by which these non-mitotic cells make this transition is not well understood. Prior studies have also suggested that skeletal stem and progenitor cells (SSPCs) localize to the surrounding periosteum and serve as a major source of marrow associated SSPCs, osteoblasts, osteocytes, and adipocytes during skeletal development. To further understand the cell transition process by which hypertrophic chondrocytes contribute to osteoblasts or other marrow associated cells, we utilized inducible and constitutive hypertrophic chondrocyte lineage tracing and reporter mouse models (Col10a1CreERT2; Rosa26fs-tdTomato and Col10a1Cre; Rosa26fs-tdTomato) in combination with a PDGFRaH2B-GFP transgenic line, single cell RNA-sequencing, bulk RNA-sequencing, immunofluorescence staining, and cell transplantation assays. Our data demonstrate that hypertrophic chondrocytes undergo a process of dedifferentiation to generate marrow associated SSPCs that serve as a primary source of osteoblasts during skeletal development. These hypertrophic chondrocyte derived SSPCs commit to a CXCL12-abundant reticular (CAR) cell phenotype during skeletal development and demonstrate unique abilities to recruit vasculature and promote bone marrow establishment, while also contributing to the adipogenic lineage.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Colony Forming Unit Assaysmentioning

confidence: 99%

Hypertrophic chondrocytes serve as a reservoir for marrow-associated skeletal stem and progenitor cells, osteoblasts, and adipocytes during skeletal development

Long

Leinroth

Liao

et al. 2022

eLife

Self Cite

View full text Add to dashboard Cite

show abstract

“…[46] As for BMSC isolation, BMSCs were isolated by flushing the bone marrow from the long bones of hind limbs. [47] Then BMSCs were cultured in high glucose DMEM medium supplemented with 20% FBS and 1% penicillin/streptomycin.…”

Section: Methodsmentioning

confidence: 99%

Hierarchically Injectable Hydrogel Sequentially Delivers AntagomiR‐467a‐3p‐Loaded and AntagomiR‐874‐5p‐Loaded Satellite‐Cell‐Targeting Bioengineered Extracellular Vesicles Attenuating Sarcopenia

Dai

Luo

Deng

et al. 2023

Adv Healthcare Materials

View full text Add to dashboard Cite

Sarcopenia is a geriatric disease characterized by reduced muscle function and mass. The capacity to self-renew and myogenesis of satellite cells (SCs) declines with age, resulting in age-related sarcopenia. MicroRNAs (miRNAs) can regulate the proliferation and myogenesis of SCs. In this study, it is identified that miR-467a-3p and miR-874-5p could respectively mediate the stemness and myogenesis of SCs by performing bioinformatics analysis. AntagomiR-467a-3p (ant-467a) and antagomiR-874-5p (ant-874) improve the stemness and myogenesis of SCs, respectively. SC-targeting extracellular vesicles (EVs) are constructed by overexpressing TSG101 on the surface of EVs isolated from bone marrow mesenchymal stem cells. Ant-467a loaded EVs (EVs-467a) and ant-874 loaded EVs (EVs-874) are prepared by transferring ant-467a and ant-874 into SC-targeting EVs. EVs-467a and EVs-874 are more effective than ant-467a and ant-874 in promoting the stemness and myogenesis of SCs. Sequentially intermuscular injection of EVs-467a and EVs-874 significantly improve sarcopenia in ovariectomy mice. The effects of multiple injections of EVs-467a and EVs-874 in the treatment of sarcopenia could be achieved by using a hierarchically injectable hydrogel to sustainedly release EVs-467a and EVs-874 in vivo. The findings provide an EV-based SC-targeting antagomiRNAs controlled release strategy as a novel therapy against sarcopenia.

show abstract

“…The previous evidence indicated that the number of CD13 + CD29 + CD44 + CD73 + CD90 + CD105 + CD151 + CD34 − BMSCs colonies developed was higher under hypoxia (3% O2) [ 92 ]. The recent study of Guo showed that PDGFRα + and LepR + BMSC subpopulations exhibited increased proliferation under 1% O2, while a 10-fold reduction in CD45 + hematopoietic cells [ 93 ]. In addition to proliferation, hypoxia also can influence specific BMSC subpopulations differentiation.…”

Section: Functional Heterogeneity Of Bmsc Subpopulations In Physiolog...mentioning

confidence: 99%

Functional Heterogeneity of Bone Marrow Mesenchymal Stem Cell Subpopulations in Physiology and Pathology

Ning

Chen

et al. 2022

IJMS

View full text Add to dashboard Cite

Bone marrow mesenchymal stem cells (BMSCs) are multi-potent cell populations and are capable of maintaining bone and body homeostasis. The stemness and potential therapeutic effect of BMSCs have been explored extensively in recent years. However, diverse cell surface antigens and complex gene expression of BMSCs have indicated that BMSCs represent heterogeneous populations, and the natural characteristics of BMSCs make it difficult to identify the specific subpopulations in pathological processes which are often obscured by bulk analysis of the total BMSCs. Meanwhile, the therapeutic effect of total BMSCs is often less effective partly due to their heterogeneity. Therefore, understanding the functional heterogeneity of the BMSC subpopulations under different physiological and pathological conditions could have major ramifications for global health. Here, we summarize the recent progress of functional heterogeneity of BMSC subpopulations in physiology and pathology. Targeting tissue-resident single BMSC subpopulation offers a potentially innovative therapeutic strategy and improves BMSC effectiveness in clinical application.

show abstract

Hypoxia depletes contaminating CD45+ hematopoietic cells from murine bone marrow stromal cell (BMSC) cultures: Methods for BMSC culture purification

Cited by 5 publications

References 44 publications

Hypertrophic chondrocytes serve as a reservoir for marrow-associated skeletal stem and progenitor cells, osteoblasts, and adipocytes during skeletal development

Hypertrophic chondrocytes serve as a reservoir for marrow-associated skeletal stem and progenitor cells, osteoblasts, and adipocytes during skeletal development

Hierarchically Injectable Hydrogel Sequentially Delivers AntagomiR‐467a‐3p‐Loaded and AntagomiR‐874‐5p‐Loaded Satellite‐Cell‐Targeting Bioengineered Extracellular Vesicles Attenuating Sarcopenia

Functional Heterogeneity of Bone Marrow Mesenchymal Stem Cell Subpopulations in Physiology and Pathology

Contact Info

Product

Resources

About