E. Andreev1, E. Kravchenko1,a, E. Zavyalova1,2, P. Eremin3, A. Rzyanina1, P. Markov3, A. Nechaev1
Keywords:
MSC, secretome, conditioned media, serum-free culture, exosome isolation, ultrafiltration
Topics:
Life Sciences
, Biology
Abstract
This study provides a comparative analysis of various components of mesenchymal stem cells (MSC) conditioned media (CM) obtained using serum-containing and serum-free culture methods, revealing significant differences in their composition and potential clinical applicability. Serum-containing CM exhibits significantly higher levels of total protein, non-vesicular RNA, exosomes, and nanoparticles compared to serum-free CM, reflecting the contribution of both the MSC secretome and residual fetal bovine serum components. Ultrafiltration-based fractionation (0.2 µm–50 kDa) allows the isolation of fraction enriched in exosomes and proteins, preserving the functionally significant components of the MSC secretome. This strategy effectively captures small vesicles and mid-sized proteins while excluding larger or smaller biomolecules, enhancing utility for targeted analyses. The presented data underscore the need for context-driven CM selection and provide information for choosing the optimal strategy for obtaining the MSC secretome balancing yield, purity, and regulatory demands in MSC research and therapy.
Acknowledgements
The authors are grateful to Nikolay Lizunov and Oleg Orelovich for the help with SEMexamination of samples, Alisher Mutali for the help with TEM examination of samples andKirill Tarasov for the data visualization. The research was funded by the Joint Institute for Nuclear Research within the frameworkof projects Nanocomposite and functional track membranes 07-5-1131-2-2024/2028, Highly sen-sitive sensors operating on the principles of molecular recognition for virus detection 07-5-1131-3-2025/2029, TARDISS 05-2-1132-1-2021/2028.
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