Evaluation of Cytocompatibility as Assessed by Genomic Stability of Canine induced Pluripotent Stem Cells Propagated on Carbon Nanotube Substrates

DOI: 10.18805/IJAR.B-4853    | Article Id: B-4853 | Page : 649-654
Citation :- Evaluation of Cytocompatibility as Assessed by Genomic Stability of Canine induced Pluripotent Stem Cells Propagated on Carbon Nanotube Substrates.Indian Journal of Animal Research.2022.(56):649-654
Tanmay Mondal, Pranay Konda, Kinshuk Das1, Kuldeep Kumar, Sadhan Bag bag658@gmail.com
Address : Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India.
Submitted Date : 29-12-2021
Accepted Date : 15-01-2022


Background: Induced pluripotent stem cells (iPSC) are a type of pluripotent stem cell derived from adult somatic cells that have been genetically reprogrammed to an embryonic stem (ES) cell-like state through the forced expression of genes and factors important for maintaining the defined properties of ES cells. So far there are very few experiments that have been able to prove that nanomaterial-based scaffold can cultivate and maintain the iPSC as an alternative to feeder-free maintenance of iPSC.
Methods: The present experiment has given us fundamental information on ex vivo canine iPSC behavior on -OH functionalized single and multi-walled carbon nanotube (CNT) scaffold. Here in we evaluated the cytocompatibility of iPSC cultured on MEF feeder, OH-SWCNT and OH-MWCNT. 
Result: We have seen very wonderful growth of ciPSC on CNTs similar to feeder. The cells were positive for alkaline phosphatase staining and expressed pluripotent markers. Cytotoxicity analysis revealed that -OH functionalized CNTs provide a milieu of low cytotoxicity. With this test we can interpret that -OH functionalized CNT can be used as xeno-free substrate to support the maintenance of iPSC in an undifferentiated state.


Canine induced pluripotent stem cells DNA ladder assay Functionalized carbon nanotube


  1. Akasaka, T., Yokoyama, A., Matsuoka, M., Hashimoto, T. and Watari, F. (2011). Maintenance of hemiround colonies and undifferentiated state of mouse induced pluripotent stem cells on carbon nanotube-coated dishes. Carbon. 49(7): 2287-2299.
  2. Amit, M. and Itskovitz Eldor, J., (2006). Feeder free culture of human embryonic stem cells. Methods in Enzymology. 420: 37- 49.
  3. Baird, A.E.G., Barsby, T. and Guest, D.J. (2015). Derivation of canine induced pluripotent stem cells. Reproduction in Domestic Animals. 50(4): 669-676.
  4. Brunner, E.W., Jurewicz, I., Heister, E., Fahimi, A., Bo, C., Sear, R.P., Donovan, P.J. and Dalton, A.B. (2014). Growth and proliferation of human embryonic stem cells on fully synthetic scaffolds based on carbon nanotubes. ACS Applied Materials and Interfaces. 6(4): 2598-2603.
  5. Das, K., Madhusoodan, A.P., Mili, B., Kumar, A., Saxena, A.C., Kumar, K., Sarkar, M., Singh, P., Srivastava, S. and Bag, S. (2017). Functionalized carbon nanotubes as suitable scaffold materials for proliferation and differentiation of canine mesenchymal stem cells. International Journal of Nano Medicine. 12: 3235.
  6. Holmes, B., Fang, X., Zarate, A., Keidar, M. and Zhang, L.G. (2016). Enhanced human bone marrow mesenchymal stem cell chondrogenic differentiation in electrospun constructs with carbon nanomaterials. Carbon. 97: 1-13.
  7. Li, X., Liu, H., Niu, X., Yu, B., Fan, Y., Feng, Q., Cui, F.Z. and Watari, F. (2012). The use of carbon nanotubes to induce osteogenic differentiation of human adipose-derived MSCs in vitro and ectopic bone formation in vivo. Biomaterials. 33(19): 4818-4827.
  8. Mahapatra, P.S., Singh, R., Kumar, K., Sahoo, N.R., Agarwal, P., Mili, B., Das, K., Sarkar, M., Bhanja, S.K., Das, B.C., Dhara, S.K. and Bag, S. (2016). Valproic acid assisted reprogramming of fibroblasts for generation of pluripotent stem cells in buffalo (Bubalus bubalis). International Journal of Developmental Biology. 61(1-2): 81-88.
  9. Marina, M. and Saavedra, H.I. (2014). Nek2 and Plk4: Prognostic markers, drivers of breast tumorigenesis and drug resistance. Frontiers in Bioscience. 19: 352.
  10. Mondal, T., Das, K., Singh, P., Natarajan, M., Manna, B., Ghosh, A., Singh, P., Saha, S.K., Dhama, K., Dutt, T. and Bag, S. (2021). Thin films of functionalized carbon nanotubes support long-term maintenance and cardio-neuronal differentiation of canine induced pluripotent stem cells. Nanomedicine: Nanotechnology, Biology and Medicine. 40(2022): 102487.
  11. Nakagawa, M., Taniguchi, Y., Senda, S., Takizawa, N., Ichisaka, T., Asano, K., Morizane, A., Doi, D., Takahashi, J., Nishizawa, M. and Yoshida, Y. (2014). A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells. Scientific Reports. 4(1): 1-7.
  12. Pan, C., Hicks, A., Guan, X., Chen, H. and Bishop, C.E. (2010). SNL fibroblast feeder layers support derivation and maintenance of human induced pluripotent stem cells. Journal of Genetics and Genomics. 37(4): 241-248.
  13. Saito, N., Haniu, H., Usui, Y., Aoki, K., Hara, K., Takanashi, S., Shimizu, M., Narita, N., Okamoto, M., Kobayashi, S. and Nomura, H. (2014). Safe clinical use of carbon nanotubes as innovative biomaterials. Chemical Reviews. 114(11): 6040-6079.
  14. Takahashi, K., Okita, K., Nakagawa, M. and Yamanaka, S. (2007). Induction of pluripotent stem cells from fibroblast  cultures. Nature Protocols. 2(12): 3081-3089.
  15. Villa Diaz, L.G., Ross, A.M., Lahann, J. and Krebsbach, P.H. (2013). Concise review: The evolution of human pluripotent stem cell culture: From feeder cells to synthetic coatings. Stem Cells. 31(1): 1-7.
  16. Wu, Y., Zhang, Y., Mishra, A., Tardif, S.D. and Hornsby, P.J. (2010). Generation of induced pluripotent stem cells from newborn marmoset skin fibroblasts. Stem Cell Research. 4(3): 180-188.

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