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Published on: July 3, 2015
Assessment of DNA damage in human bone marrow cells and multipotent mesenchymal stromal cells
V A Nikitina1, A I Chausheva, A K Zhanataev
1Medical Genetic Research Center, Russian Academy of Medical Sciences, Moscow, Russia. vanikitina@mail.ru
Abstract:
We carried out a comparative analysis of DNA damage (percentage of DNA in comet tail) and frequencies of comets in apoptotic cells in BM samples and cultures of BM multipotent mesenchymal stromal cells at different terms of culturing (passages 3-11). The levels of DNA damage in mesenchymal stromal cells remained unchanged during culturing (3.5 ± 0.9 and 4.4 ± 1.2%) and did not differ from those in BM cells (3.6 ± 0.8%). In BM samples, 10-28% atypical cells with high level of DNA damage were detected. In mesenchymal stromal cells, 2.8 ± 0.9 and 3.6 ± 1.8% apoptotic cells were detected at early and late passages, respectively.
Insights
Mesenchymal stromal cells (MSCs) cultured in vitro show low DNA damage and apoptosis, similar to bone marrow (BM) cells. This indicates MSCs are safe for research and potential therapies.
Area of Science:
- Cell Biology
- Genetics
- Biomedical Research
Background:
- Bone marrow (BM) multipotent mesenchymal stromal cells (MSCs) are crucial for regenerative medicine.
- Assessing DNA damage and apoptosis in cultured MSCs is vital for safety and efficacy.
- Understanding MSC behavior during in vitro culturing is essential for therapeutic applications.
Purpose of the Study:
- To comparatively analyze DNA damage and apoptosis in BM samples versus cultured MSCs.
- To evaluate the stability of DNA integrity and apoptotic rates in MSCs across multiple passages.
- To determine the safety profile of MSCs for potential clinical use.
Main Methods:
- Comet assay to quantify DNA damage (percentage of DNA in comet tail).
- Analysis of comet frequencies in apoptotic cells within BM samples and cultured MSCs.
- Culturing of BM MSCs across different passages (3-11).
Main Results:
- DNA damage levels in cultured MSCs remained stable across passages (3.5 ± 0.9% to 4.4 ± 1.2%) and were comparable to BM cells (3.6 ± 0.8%).
- BM samples exhibited 10-28% atypical cells with significant DNA damage.
- Apoptotic cells in cultured MSCs were low, at 2.8 ± 0.9% (early passages) and 3.6 ± 1.8% (late passages).
Conclusions:
- Cultured MSCs demonstrate minimal DNA damage and low apoptosis rates, mirroring native BM cells.
- The in vitro culturing process does not induce significant genotoxicity or increase apoptosis in MSCs.
- These findings support the safety and suitability of MSCs for further research and therapeutic development.
