Advertisement

Chemical Reprogramming of Somatic Cells in Neural Direction: Myth or Reality?

  • E. M. SamoilovaEmail author
  • V. A. Revkova
  • O. I. Brovkina
  • V. A. Kalsin
  • P. A. Melnikov
  • M. A. Konoplyannikov
  • K. R. Galimov
  • A. G. Nikitin
  • A. V. Troitskiy
  • V. P. Baklaushev
Article

In in vitro experiments on cultures of human multipotent stem cells from the human bone

arrow and dental pulp, we studied direct reprogramming towards neuro-glial lineage cells using a cocktail of small molecules. Reprogramming by the previously published protocol (with a cocktail containing β-mercaptoethanol, LIF, VPA, CHIR99021, and RepSox) and by the optimized protocol (VPA, RG108, А83-01, dorsomorphin, thiazovivin, CHIR99021, forskolin, and Isx9) allows obtaining cells with immunophenotypic and genetic signs of neural stem cells. However, neither the former, nor the optimized protocols allowed preparing neural progenitors capable of adequate terminal differentiation from both bone marrow-derived mesenchymal stem cells and nestin-positive neural crest-derived mesenchymal stem cells. Real-time PCR demonstrated the expression of some neurogenesis markers, but neural stem cell-specific expression pattern was not observed. The findings lead us to a conclusion that reprogramming with small molecules without additional factors modifying gene expression does not allow reproducible production of human neural stem cell-like progenitors that can be used as the source of neural tissue for the regenerative therapy.

Key Words

chemical reprogramming small molecules CHIR99021 Isx9 neural stem cells 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bettio LEB, Gil-Mohapel J, Patten AR, O’Rourke NF, Hanley RP, Gopalakrishnan K, Wulff JE, Christie BR. Effects of Isx-9 and stress on adult hippocampal neurogenesis: Experimental considerations and future perspectives. Neurogenesis (Austin). 2017;4(1). ID e1317692. doi:  https://doi.org/10.1080/23262133.2017.1317692
  2. 2.
    Blanchard JW, Eade KT, Szűcs A, Lo Sardo V, Tsunemoto RK, Williams D, Sanna PP, Baldwin KK. Selective conversion of fibroblasts into peripheral sensory neurons. Nat. Neurosci. 2015;18(1):25-35.CrossRefGoogle Scholar
  3. 3.
    Biswas D, Jiang P. Chemically induced reprogramming of somatic cells to pluripotent stem cells and neural cells. Int. J. Mol. Sci. 2016;17(2). ID 226. doi:  https://doi.org/10.3390/ijms17020226
  4. 4.
    Caiazzo M, Dell’Anno MT, Dvoretskova E, Lazarevic D, Taverna S, Leo D, Sotnikova TD, Menegon A, Roncaglia P, Colciago G, Russo G, Carninci P, Pezzoli G, Gainetdinov RR, Gustincich S, Dityatev A, Broccoli V. Direct generation of functional dopaminergic neurons from mouse and human fibroblasts. Nature. 2011;476:224-227.CrossRefGoogle Scholar
  5. 5.
    Cheng L, Hu W, Qiu B, Zhao J, Yu Y, Guan W, Wang M, Yang W, Pei G. Generation of neural progenitor cells by chemical cocktails and hypoxia. Cell Res. 2014;24(6):665-679.CrossRefGoogle Scholar
  6. 6.
    Fu Y, Huang C, Xu X, Gu H, Ye Y, Jiang C, Qiu Z, Xie X. Direct reprogramming of mouse fibroblasts into cardiomyocytes with chemical cocktails. Cell Res. 2015;25(9):1013-1024.CrossRefGoogle Scholar
  7. 7.
    Fusaki N, Ban H, Nishiyama A, Saeki K, Hasegawa M. Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proc. Jpn. Acad. Ser. B Phys. Biol. Sci. 2009;85(8):348-362.CrossRefGoogle Scholar
  8. 8.
    Hou P, Li Y, Zhang X, Liu C, Guan J, Li H, Zhao T, Ye J, Yang W, Liu K, Ge J, Xu J, Zhang Q, Zhao Y, Deng H. Pluripotent stem cells induced from mouse somatic cells by small-molecule compounds. Science. 2013;341:651-654.CrossRefGoogle Scholar
  9. 9.
    Kelaini S, Cochrane A, Margariti A. Direct reprogramming of adult cells: avoiding the pluripotent state. Stem Cells Cloning. 2014;7:19-29.Google Scholar
  10. 10.
    Kim SM, Flaßkamp H, Hermann A, Araúzo-Bravo MJ, Lee SC, Lee SH, Seo EH, Lee SH, Storch A, Lee HT, Schöler HR, Tapia N, Han DW. Direct conversion of mouse fibroblasts into induced neural stem cells. Nat. Protoc. 2014;9(4):871-881.CrossRefGoogle Scholar
  11. 11.
    Li H, Radford JC, Ragusa MJ, Shea KL, McKercher SR, Zaremba JD, Soussou W, Nie Z, Kang YJ, Nakanishi N, Okamoto S, Roberts AJ, Schwarz JJ, Lipton SA. Transcription factor MEF2C influences neural stem/progenitor cell differentiation and maturation in vivo. Proc. Natl Acad. Sci. USA. 2008;105(27):9397-9402.CrossRefGoogle Scholar
  12. 12.
    Li X, Zuo X, Jing J, Ma Y, Wang J, Liu D, Zhu J, Du X, Xiong L, Du Y, Xu J, Xiao X, Wang J, Chai Z, Zhao Y, Deng H. Small-molecule-driven direct reprogramming of mouse fibroblasts into functional neuros. Cell Stem Cell. 2015;17(2):195-203.CrossRefGoogle Scholar
  13. 13.
    Liu JA, Cheung M. Neural crest stem cells and their potential therapeutic applications. Dev. Biol. 2016;419(2):199-216.CrossRefGoogle Scholar
  14. 14.
    Meraviglia V, Zanon A, Lavdas AA, Schwienbacher C, Silipigni R, Di Segni M, Chen H.S, Pramstaller PP, Hicks AA, Rossini A. Generation of induced pluripotent stem cells from frozen buffy coats using non-integrating episomal plasmids. J. Vis. Exp. 2015. N 100. ID e52885. doi:  https://doi.org/10.3791/52885
  15. 15.
    Nagoshi N, Khazaei M, Ahlfors JE, Ahuja CS, Nori S, Wang J, Shibata S, Fehlings MG. Human spinal oligodendrogenic neural progenitor cells promote functional recovery after spinal cord injury by axonal remyelination and tissue sparing. Stem Cells Transl. Med. 2018;7(11):806-818.CrossRefGoogle Scholar
  16. 16.
    Potthoff MJ, Olson EN. MEF2: a central regulator of diverse developmental programs. Development. 2007;134(23):4131-4140.CrossRefGoogle Scholar
  17. 17.
    Poulos J. The limited application of stem cells in medicine: a review. Stem Cell Res. Ther. 2018;9(1). doi:  https://doi.org/10.1186/s13287-017-0735-7
  18. 18.
    Qin H, Zhao A, Fu X. Small molecules for reprogramming and transdifferentiation. Cell Mol. Life Sci. 2017 Vol. 74(19):3553-3575.CrossRefGoogle Scholar
  19. 19.
    Rikhtegar R, Pezeshkian M, Dolati S, Safaie N, Afrasiabi Rad A, Mahdipour M, Nouri M, Jodati A. R, Yousefi M. Stem cells as therapy for heart disease: iPSCs, ESCs, CSCs, and skeletal myoblasts. Biomed. Pharmacother. 2018;109:304-313.CrossRefGoogle Scholar
  20. 20.
    Samoilova EM, Kalsin VA, Kushnir NM, Chistyakov DA, Troitskiy AV, Baklaushev VP. Adult neural stem cells: basic research and production strategies for neurorestorative therapy. Stem Cells Int. 2018;2018. ID 4835491. doi:  https://doi.org/10.1155/2018/4835491
  21. 21.
    Son EY, Ichida JK, Wainger BJ, Toma JS, Rafuse VF, Woolf CJ, Eggan K. Conversion of mouse and human fibroblasts into functional spinal motor neurons. Cell Stem Cell. 2011;9(3):205-218.CrossRefGoogle Scholar
  22. 22.
    Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131(5):861-872.CrossRefGoogle Scholar
  23. 23.
    Vierbuchen T, Wernig M. Molecular roadblocks for cellular reprogramming. Mol. Cell. 2012;47(6):827-838.CrossRefGoogle Scholar
  24. 24.
    Warren L, Ni Y, Wang J, Guo X. Feeder-free derivation of human induced pluripotent stem cells with messenger RNA. Sci. Rep. 2012;2. ID 657. doi:  https://doi.org/10.1038/srep00657
  25. 25.
    Yamamoto A, Sakai K, Matsubara K, Kano F, Ueda M. Multifaceted neuro-regenerative activities of human dental pulp stem cells for functional recovery after spinal cord injury. Neurosci. Res. 2014;78:16-20.CrossRefGoogle Scholar
  26. 26.
    Zhou H, Wu S, Joo JY, Zhu S, Han DW, Lin T, Trauger S, Bien G, Yao S, Zhu Y, Siuzdak G, Schöler H. R, Duan L, Ding S. Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell. 2009;4(5):381-384.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • E. M. Samoilova
    • 1
    Email author
  • V. A. Revkova
    • 1
  • O. I. Brovkina
    • 1
  • V. A. Kalsin
    • 1
  • P. A. Melnikov
    • 1
    • 2
  • M. A. Konoplyannikov
    • 1
    • 3
  • K. R. Galimov
    • 1
  • A. G. Nikitin
    • 1
  • A. V. Troitskiy
    • 1
  • V. P. Baklaushev
    • 1
  1. 1.Federal Research Clinical Center of Specialized Medical Care, Federal Medical-Biological Agency of RussiaMoscowRussia
  2. 2.Department of Fundamental and Applied Neurobiology, V. P. Serbsky Federal Medical Research Center for Psychiatry and Narcology, Ministry of Health of the Russian FederationMoscowRussia
  3. 3.Institute of Regenerative Medicine, I. M. Sechenov First Moscow State Medical University (Sechenov University)MoscowRussia

Personalised recommendations