ORIGINAL RESEARCH
Survival of human cells in tissue-engineered constructs stored at room temperature
1 Koltzov Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia
2 Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of the Federal Medical Biological Agency, Moscow, Russia
Correspondence should be addressed: Olga S. Rogovaya
Vavilova, 26, Moscow, 119334, Russia; ur.xednay@f62ayavogor
Funding: the study was supported by the Ministry of Science and Higher Education of the Russian Federation, Agreement № 075-15-2021-1063 of 28.09.2021.
Author contribution: Rogovaya OS, Eremeev AV — experimental procedure, data analysis; Alpeeva EV — data interpretation, literature review; Ruchko ES — experimental procedure; Vorotelyak EA — study planning.
Compliance with ethical standards: the study was approved by the Ethics Committee of the Koltzov Institute of Developmental Biology, RAS (protocol № 51 of 09 September 2021) and conducted in accordance with the principles of the WMA Declaration of Helsinki and its subsequent revisions.
Tissue-engineered constructs (TECs), the dermal equivalent (DE) and the skin equivalent (SE), are allogenic equivalents of the skin and derm used to treat critical skin loss. Selection of storage conditions that contribute to longer shelf life, thereby expanding the possibilities of logistics and use, is one of the major issues related to the TECs development. The study was aimed to determine the shelf life of the DE and SE TECs stored in normal saline at room temperature by assessing morphology and viability of the cells on their surface, along with the levels of endothelial growth factor (VEGF) secreted by these cells. Using the MTT assay and staining with vital dye, we discovered the following: when TECs of both types were stored in normal saline, the cells viability and metabolic activity decreased by more than 50% by days 3–4 of storage. Furthermore, these decreased faster in DEs than in SEs. Morphology of the cells isolated from DEs and SEs after the 3-day storage remained unchanged. Mesenchymal stem cells on the surface of TECs kept producing VEGF after TECs culture medium was changed for saline solution (confirmed by immunofluorescence assay), which could indicate that the cells retained essential secretory activity.
Keywords: tissue-engineered construct, TEC, skin equivalent, mild hypothermia, keratinocytes, MSCs