Development of secretome‑based strategies to improve cell culture protocols in tissue engineering

This study was supported by the Spanish Plan Nacional de Investigación Científica, Desarrollo e Innovación Tec- nológica (I + D + I) of the Spanish Ministry of Science and Innovation (Instituto de Salud Carlos III), grants FIS PI18/0331, FIS PI21/0980, FIS PI18/0332, FIS PI20/0317, ICI19/00024 and ICI21/00010, and by grants PE-0395- 2019 and PI-0442-2019 from the Consejería de Salud y Familias, Junta de Andalucía, Spain. Additional support was provided through grant B-CTS-450-UGR20 (proyectos de I + D + i en el marco del Programa Operativo FEDER Andalucía 2014–2020, University of Granada and Consejería de Transformación Económica, Industria, Conocimiento y Universidades), and cofinancing was provided from the European Regional Development Fund (ERDF) through the “Una manera de hacer Europa” program.The online version contains supplementary material available at https://doi. org/10. 1038/s41598-022-14115-yAdvances in skin tissue engineering have promoted the development of artificial skin substitutes to treat large burns and other major skin loss conditions. However, one of the main drawbacks to bioengineered skin is the need to obtain a large amount of viable epithelial cells in short periods of time, making the skin biofabrication process challenging and slow. Enhancing skin epithelial cell cultures by using mesenchymal stem cells secretome can favor the scalability of manufacturing processes for bioengineered skin. The effects of three different types of secretome derived from human mesenchymal stem cells, e.g. hADSC‑s (adipose cells), hDPSC‑s (dental pulp) and hWJSC‑s (umbilical cord), were evaluated on cultured skin epithelial cells during 24, 48, 72 and 120 h to determine the potential of this product to enhance cell proliferation and improve biofabrication strategies for tissue engineering. Then, secretomes were applied in vivo in preliminary analyses carried out on Wistar rats. Results showed that the use of secretomes derived from mesenchymal stem cells enhanced currently available cell culture protocols. Secretome was associated with increased viability, proliferation and migration of human skin epithelial cells, with hDPSC‑s and hWJSC‑s yielding greater inductive effects than hADSC‑s. Animals treated with hWJSC‑s and especially, hDPSC‑s tended to show enhanced wound healing in vivo with no detectable side effects. Mesenchymal stem cells derived secretomes could be considered as a promising approach to cell‑free therapy able to improve skin wound healing and regeneration.Spanish Plan Nacional de Investigacion Cientifica, Desarrollo e Innovacion Tecnologica (I + D + I) of the Spanish Ministry of Science and Innovation (Instituto de Salud Carlos III) FIS PI18/0331, FIS PI21/0980, FIS PI18/0332, FIS PI20/0317, ICI19/00024, ICI21/00010Junta de Andalucia PE-0395-2019, PI-0442-2019Proyectos de I + D + i en el marco del Programa Operativo FEDER Andalucia , University of Granada and Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades B-CTS-450-UGR20European Commissio