Effect of Aging and 5-Fluorouracil Treatment on Bone Marrow Stem Cell Dynamics


Lifelong homeostasis of bone marrow is maintained by the resident stem cells that include the quiescent very small embryonic-like stem cells (VSELs) and lineage restricted, tissue committed ‘progenitors’ hematopoietic stem cells (HSCs). Niche providing mesenchymal stromal cells (MSCs) regulate the function of VSELs/HSCs by providing crucial paracrine support. Any dysfunction of stem cells and/or their niche leads to disease state. The stem cells biology gets affected with age leading to a myeloid bias in differentiation of HSCs and increased incidence of myeloid leukemia. Present study was undertaken to enumerate VSELs, HSCs and MSCs and evaluate their response on D4 and D10 after chemotherapy with 5-Fluorouracil (5-FU) in young and aged mouse bone marrow. Stem cells were activated in response to 5-FU induced stress in an attempt to restore homeostasis. Although absolute numbers of VSELs and HSCs did not differ much between young and aged mice, their tendency to proliferate was higher on D4 in aged mice. However, ability to revert back to basal numbers and their differentiation was affected on D10 in aged marrow. Stem cells from aged bone marrow showed greater ability to form CFUs on D10 after 5-FU treatment. CD44 positive aged MSCs also showed increased proliferation on D10. Transplanting MSCs from young mice in 5-FU treated aged marrow helped improve hematopoiesis. The results suggest that no significant intrinsic changes occur as proliferative ability of stem cells remains unaffected but the niche gets affected with age leading to excessive self-renewal and compromised differentiation. This may explain increased incidence of leukemia with age.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8


  1. 1.

    Bhartiya, D., Patel, H., Ganguly, R., Shaikh, A., Shukla, Y., Sharma, D., & Singh, P. (2018). Novel insights into adult and cancer stem cell biology. Stem Cells and Development, 27(22), 1527–1539.

    Article  Google Scholar 

  2. 2.

    Konieczny, J., & Arranz, L. (2018). Updates on old and weary Haematopoiesis. International Journal of Molecular Sciences, 19(9), E2567.

    Article  Google Scholar 

  3. 3.

    Gustafsson, K., & Scadden, D. T. (2017). Written in bone: Young bone makes young blood. The EMBO Journal, 36(7), 831–833.

    CAS  Article  Google Scholar 

  4. 4.

    Pearce, D. J., Anjos-Afonso, F., Ridler, C. M., Eddaoudi, A., & Bonnet, D. (2007). Age-dependent increase in side population distribution within hematopoiesis: Implications for our understanding of the mechanism of aging. Stem Cells, 25, 828–835.

    CAS  Article  Google Scholar 

  5. 5.

    Rossi, D. J., Bryder, D., Zahn, J. M., Ahlenius, H., Sonu, R., Wagers, A. J., & Weissman, I. L. (2005). Cell intrinsic alterations underlie hematopoietic stem cell aging. Proceedings of the National Academy of Sciences of the United States of America, 102(26), 9194–9199.

    CAS  Article  Google Scholar 

  6. 6.

    Sudo, K., Ema, H., Morita, Y., & Nakauchi, H. (2000). Age associated characteristics of murine hematopoietic stem cells. The Journal of Experimental Medicine, 192(9), 1273–1280.

    CAS  Article  Google Scholar 

  7. 7.

    Ho, Y. H., & Méndez-Ferrer, S. (2020). Microenvironmental contributions to hematopoietic stem cell aging. Haematologica, 105(1), 38–46.

    Article  Google Scholar 

  8. 8.

    Chambers, S. M., Shaw, C. A., Gatza, C., Fisk, C. J., Donehower, L. A., & Goodell, M. A. (2007). Aging hematopoietic stem cells decline in function and exhibit epigenetic dysregulation. PLoS Biology, 5, e201.

    Article  Google Scholar 

  9. 9.

    Ratajczak, M. Z., Ratajczak, J., & Kucia, M. (2019). Very small embryonic-like stem cells (VSELs). Circulation Research, 124(2), 208–210.

    CAS  Article  Google Scholar 

  10. 10.

    Ganguly, R., Metkari, S., & Bhartiya, D. (2018). Dynamics of bone marrow VSELs and HSCs in response to treatment with gonadotropin and steroid hormones, during pregnancy and evidence to support their asymmetric/symmetric cell divisions. Stem Cell Reviews and Reports, 14(1), 110–124.

    CAS  Article  Google Scholar 

  11. 11.

    Sovalat, H., Scrofani, M., Eidenschenk, A., et al. (2015). Human very small embryonic-like stem cells are present in normal peripheral blood of young, middle-aged, and aged subjects. Stem Cells International, 2016, 7651645.

    PubMed  PubMed Central  Google Scholar 

  12. 12.

    Mejia-Ramirez, E., & Florian, M. C. (2020). Understanding intrinsic hematopoietic stem cell aging. Haematologica, 105(1), 22–37.

    Article  Google Scholar 

  13. 13.

    Shaikh, A., Bhartiya, D., Kapoor, S., & Nimkar, H. (2016). Delineating the effects of 5-fluorouracil and follicle-stimulating hormone on mouse bone marrow stem/progenitor cells. Stem Cell Research & Therapy, 7(1), 59.

    Article  Google Scholar 

  14. 14.

    Soleimani, M., & Nadri, S. (2009). A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nature Protocols, 4(1), 102–106.

    CAS  Article  Google Scholar 

  15. 15.

    Anand, S., Bhartiya, D., Sriraman, K., & Mallick, A. (2016). Underlying mechanisms that restore spermatogenesis on transplanting healthy niche cells in busulphan treated mouse testis. Stem Cell Reviews and Reports, 12(6), 682–697.

    CAS  Article  Google Scholar 

  16. 16.

    Pang, W. W., Price, E. A., Sahoo, D., Beerman, I., Maloney, W. J., Rossi, D. J., Schrier, S. L., & Weissman, I. L. (2011). Human bone marrow hematopoietic stem cells are increased in frequency and myeloid-biased with age. Proceedings of the National Academy of Sciences of the United States of America, 108(50), 20012–20017.

    CAS  Article  Google Scholar 

  17. 17.

    Fumagalli, M., Rossiello, F., Clerici, M., Barozzi, S., Cittaro, D., Kaplunov, J. M., Bucci, G., Dobreva, M., Matti, V., Beausejour, C. M., Herbig, U., Longhese, M. P., & d’Adda di Fagagna, F. (2012). Telomeric DNA damage is irreparable and causes persistent DNA-damage-response activation. Nature Cell Biology, 14(4), 355–365.

    CAS  Article  Google Scholar 

  18. 18.

    Crane, G. M., Jeffery, E., & Morrison, S. J. (2017). Adult haematopoietic stem cell niches. Nature Reviews. Immunology, 17(9), 573–590.

    CAS  Article  Google Scholar 

  19. 19.

    Morrison, S. J., & Scadden, D. T. (2014). The bone marrow niche for haematopoietic stem cells. Nature, 505(7483), 327–334.

    CAS  Article  Google Scholar 

  20. 20.

    Neri, S., & Borzì, R. M. (2020). Molecular mechanisms contributing to Mesenchymal stromal cell aging. Biomolecules, 10(2), E340.

    Article  Google Scholar 

  21. 21.

    de Haan, G., & Lazare, S. S. (2018). Aging of hematopoietic stem cells. Blood, 131(5), 479–487.

    Article  Google Scholar 

  22. 22.

    Budgude, P., Kale, V., & Vaidya, A. (2020). Mesenchymal stromal cell-derived extracellular vesicles as cell-free biologics for the ex vivo expansion of hematopoietic stem cells. Cell Biology International, 44(5), 1078–1102.

    CAS  Article  Google Scholar 

  23. 23.

    Wilkinson, A. C., Ishida, R., Nakauchi, H., & Yamazaki, S. (2020). Long-term ex vivo expansion of mouse hematopoietic stem cells. Nature Protocols, 15(2), 628–648.

    CAS  Article  Google Scholar 

  24. 24.

    Hao, T., Li-Talley, M., Buck, A., & Chen, W. (2019). An emerging trend of rapid increase of leukemia but not all cancers in the aging population in the United States. Scientific Reports, 9(1), 12070.

    Article  Google Scholar 

  25. 25.

    Festa (2018). [online] Available at: https://www.healthgrades.com/right-care/leukemia/leukemia-risk-by-age-group

  26. 26.

    Balducci, L., Aapro, M. [online] Available at: http://eknygos. Lsmuni.lt/springer/53/01–15.

  27. 27.

    Ratajczak, M. Z., Bujko, K., Mack, A., Kucia, M., & Ratajczak, J. (2018). Cancer from the perspective of stem cells and misappropriated tissue regeneration mechanisms. Leukemia, 32(12), 2519–2526.

    CAS  Article  Google Scholar 

  28. 28.

    Ratajczak, M. Z., Shin, D. M., Ratajczak, J., Kucia, M., & Bartke, A. (2010). A novel insight into aging: Are there pluripotent very small embryonic-like stem cells (VSELs) in adult tissues overtime depleted in an Igf-1-dependent manner? Aging (Albany NY), 2(11), 875–883.

    CAS  Article  Google Scholar 

  29. 29.

    Kurkure, P., Prasad, M., Dhamankar, V., et al. (2015). Very small embryonic-like stem cells (VSELs) detected in azoospermic testicular biopsies of adult survivors of childhood cancer. Reproductive Biology and Endocrinology, 13, 122.

    Article  Google Scholar 

  30. 30.

    Bhartiya, D., & Anand, S. (2017). Effects of oncotherapy on testicular stem cells and niche. Molecular Human Reproduction, 23(9), 654–655.

    CAS  Article  Google Scholar 

Download references


The help from Dr. Mukherjee, Ms. Gayatri and Ms. Sushma in flow cytometry studies and from Ms. Reshma and Ms. Shobha in confocal studies is also acknowledged. We thank Dr. Ekta Khattar (NMIMS Sunandan Divatia School of Science) for providing the antibodies for TRF2 and γH2AX. We also thank Indian Council of Medical Research and Department of Science and Technology, Government of India, New Delhi for providing financial support for the study and supporting RG with Women Scientist Fellowship under Scheme-A (SR/WOSA/LS-1318/2014).


Financial Support for the study was provided by Indian Council of Medical Research and Department of Science and Technology, Government of India, New Delhi, India.

Author information




RG: Study design, Collection of data, Data analysis and interpretation, Manuscript writing, Final approval of manuscript; SA: Collection of data, Data analysis and interpretation, Manuscript writing, Final approval of manuscript; SM: Experimental help in animal transplantation experiments, Final approval of manuscript; DB: Study design, Data analysis and interpretation, Manuscript writing, Final approval of manuscript.

Corresponding author

Correspondence to Deepa Bhartiya.

Ethics declarations

The study was approved by Institute Animal Ethics Committee and we have approval from NIRRH to submit this manuscript.


Authors declare no conflict of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ganguly, R., Anand, S., Metkari, S. et al. Effect of Aging and 5-Fluorouracil Treatment on Bone Marrow Stem Cell Dynamics. Stem Cell Rev and Rep (2020). https://doi.org/10.1007/s12015-020-09998-1

Download citation


  • Stem cells
  • Niche
  • HSCs
  • VSELs
  • MSCs
  • Proliferation
  • Differentiation