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Mitomycin C Treatment of Stromal Layers Enhances the Support of In Vitro Hematopoiesis in Co-Culture Systems

  • MOLECULAR CELL BIOLOGY
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Abstract

A study was made of the effect that mitomycin C (MitC) treatment of stromal layers of NIH 3T3 cells expressing Jagged1, a ligand of the Notch receptor, exerts on the growth of hematopoietic Lin(–) mouse bone marrow cells in a co-culture system. MitC treatment of stromal cells significantly increased the number of hematopoietic cells and the frequency of colony-forming cells in stromal co-cultures. Transcriptome analysis of control and MitC-treated stromal cell samples was performed by differential RNA sequencing, and genes downregulated by MitC treatment were predominantly associated with the control of cell proliferation, the cell cycle, chromosome segregation, and DNA metabolism. Induction of key hematopoietic cytokines by MitC was not detected by the transcriptome analysis and was therefore not a main factor in the activation of hematopoiesis on the treated stroma. At the same time, the set of the genes most strongly upregulated by MitC treatment is enriched in the genes for cytokines, growth factors, and cell surface proteins, which presumably contribute to enhanced hematopoiesis support on the MitC-treated stroma. Products of some of these genes have been implicated in expansion of hematopoietic stem/progenitor cells in vitro or in vivo.

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ACKNOWLEDGMENTS

We are grateful to Genoanalitika (Moscow) for the skilful transcriptome analysis and primary bioinformatics data processing.

Funding

This work was supported by the Russian Science Foundation (project no. 18-14-00300).

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Authors and Affiliations

  1. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991, Moscow, Russia

    O. F. Kandarakov, Yu. V. Kravatsky, N. S. Polyakova, A. V. Bruter, E. G. Gordeeva & A. V. Belyavsky

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  1. O. F. Kandarakov
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  2. Yu. V. Kravatsky
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  3. N. S. Polyakova
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  4. A. V. Bruter
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  5. E. G. Gordeeva
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  6. A. V. Belyavsky
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Corresponding author

Correspondence to A. V. Belyavsky.

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The authors declare that they have no conflict of interest.

This work does not contain any studies involving animals or human subjects performed by any of the authors.

Additional information

Translated by T. Tkacheva

Abbreviations: MitC, mitomycin C; LNGFR, low-affinity nerve growth factor receptor; 3T3-J/L, NIH 3T3-Jagged1-LNGFR; HSPC, hematopoietic stem/progenitor cell; RNA-Seq, high-throughput parallel RNA sequencing.

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Kandarakov, O.F., Kravatsky, Y.V., Polyakova, N.S. et al. Mitomycin C Treatment of Stromal Layers Enhances the Support of In Vitro Hematopoiesis in Co-Culture Systems. Mol Biol 55, 109–120 (2021). https://doi.org/10.1134/S0026893321010088

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  • DOI: https://doi.org/10.1134/S0026893321010088

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