Bone marrow stromal cells (BMSCs CD45 ‐ /CD44 + /CD73 + /CD90 + ) isolated from osteoporotic mice SAM/P6 as a novel model for osteoporosis investigation
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Abstract Available therapies aimed at treating age‐related osteoporosis are still insufficient. Therefore, designing reliable in vitro model for the analysis of molecular mechanisms underlying senile osteoporosis is highly required. We have isolated and characterized progenitor cells isolated from bone marrow (BMSCs) of osteoporotic mice strain SAM/P6 (BMSC SAM/P6 ). The cytophysiology of BMSC SAM/P6 was for the first time compared with BMSCs isolated from healthy BALB/c mice (BMSC BALB/c ). Characterization of the cells included evaluation of their multipotency, morphology and determination of specific phenotype. Viability of BMSCs cultures was determined in reference to apoptosis profile, metabolic activity, oxidative stress, mitochondrial membrane potential and caspase activation. Additionally, expression of relevant biomarkers was determined with RT‐qPCR. Obtained results indicated that BMSC SAM/P6 and BMSC BALB/c show the typical phenotype of mesenchymal stromal cells (CD44+, CD73+, CD90+) and do not express CD45. Further, BMSC SAM/P6 were characterized by deteriorated multipotency, decreased metabolic activity and increased apoptosis occurrence, accompanied by elevated oxidative stress and mitochondria depolarisation. The transcriptome analyses showed that BMSC SAM/P6 are distinguished by lowered expression of molecules crucial for proper osteogenesis, including Coll‐1 , Opg and Opn . However, the expression of Trap , DANCR1 and miR‐124‐3p was significantly up‐regulated. Obtained results show that BMSC SAM/P6 present features of progenitor cells with disturbed metabolism and could serve as appropriate model for in vitro investigation of age‐dependent osteoporosis.
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BMSCs were identified by flow cytometry using CD44 and CD90 as positive markers and CD34 and CD45 as negative markers in accordance with ISCT criteria, and their tri-lineage differentiation potential was confirmed.
“The cell purity was assessed based on the expression levels of CD44 (BioLegend, USA) and CD90 (BioLegend, USA) as positive markers, and CD34 (BioLegend, USA) and CD45 (BioLegend, USA) as negative markers, in accordance with the criteria established by the International Society for Cellular Therapy (ISCT) [45].”
From Hydrogel scaffold encapsulating MSC-Exos and ZIF-8 promotes bone regeneration via coordinating osteogenesis and immunomodulation (2025) · cites this paper as [45]
The Results (Characterization of isolated BMSCs; page unavailable) report CD44 and CD90 expression, lack of CD45 expression, and differentiation under chondrogenic, osteogenic, and adipogenic conditions. The Introduction lists CD34 negativity among ISCT criteria, but the paper does not report testing its cells for CD34. The Methods describe immunocytochemical staining and confocal microscopy, not flow cytometry, for the measured markers. (LLM, conf=0.88)
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- Authors
- Sikora, M., Śmieszek, A., & Marycz, K.
- Affiliations
- Wrocław University of Environmental and Life Sciences (PL)
- Corresponding author
- Marycz, Krzysztof
- Venue
- Journal of Cellular and Molecular Medicine
- Publisher
- Wiley
- Published
- 2021-06-01
- Volume
- 25
- Issue
- 14
- Pages
- 6634-6651
- ISSN
- 1582-1838 (ISSN-L)
- 1582-4934
- PMC
- PMC8278098
- OpenAlex
- W3165711336
- Cited by
- 35
- References
- 75
- Citations by year
- 2026: 5 · 2025: 7 · 2024: 10 · 2023: 8 · 2022: 3 · 2021: 2
- FWCI
- 3.37
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Journal of Cellular and Molecular Medicine (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Narodowe Centrum Nauki (UMO 2017/26/M/NZ5/01184)
- Topics
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
- Health Sciences › Medicine › Orthopedics and Sports Medicine › Bone health and osteoporosis research
- Keywords
- mesenchymal stem cells, senile osteoporosis, apoptosis, oxidative stress, mitochondrial depolarization, cellular metabolism, bone formation, miR-124-3p, traps, osteoprotegerin, osteopontin, type I collagen
- MeSH terms
- Animals, Cell Differentiation, Humans, Mice, Inbred BALB C, Osteoblasts, Osteogenesis, Osteoporosis, Stem Cells, 5'-Nucleotidase, Leukocyte Common Antigens, Thy-1 Antigens, Hyaluronan Receptors, Mice, Mesenchymal Stem Cells
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