Study on viability and chondrogenic differentiation of cryopreserved adipose tissue-derived mesenchymal stromal cells for future use in regenerative medicine.

Adipose-derived mesenchymal stromal cells are promising as a regenerative therapy tool for defective tissues in mesenchymal lineage, including fat, bone, cartilage, and blood vessels. In potential future clinical applications, adipose-derived stem cell cryopreservation is an essential fundamental technology. The aim of this study is to define an adequate protocol for the cryopreservation of adipose-derived mesenchymal stromal cells, by comparing various protocols so as to determine the effects of cryopreservation on viability and chondrogenic differentiation potential of adipose-derived stem cells upon freeze-thawing of AT-MSCs colonies cryopreserved with standard and modified protocols, using flow cytometry and confocal microscopy. The study concludes that adipose-derived mesenchymal stromal cells could be long-term cryopreserved without any loss of their proliferative or differentiation potential.

Effect of ascorbic and chondrogenic derived decellularized extracellular matrix from mesenchymal stem cells on their proliferation, viability and differentiation.

BACKGROUND: The development and application of biomaterials to promote stem cell proliferation and differentiation has undergone major expansion over the last few years. Decellularized stem cell matrix (DSCMs) represent bioactive and biocompatible materials which achieve similar characteristics of native extracellular matrix. DSCMs have given promising outcomes in generating novel cell culture substrates mimicking specific niche microenvironments in tissue engineering. AIMS: This research aims at producing two different DSCMs obtained from adipose derived mesenchymal stem cells and bone marrow mesenchymal stem cells, characterize them and evaluate the DSCMs bioactivity on mesenchymal stem cells. METHODS: DSCMs were produced using ascorbic or chondrogenic medium, which were then used as a scaffold for adipose derived mesenchymal stem cells and bone marrow mesenchymal stem cells, respectively. The biological characteristics of both types of DSCMs, including cell attachment, morphology, proliferation, viability, and chondrogenic and osteogenic differentiation were evaluated and compared. RESULTS: Differences between ascorbic derived-DSCMs and chondrogenic derived DSCMs were found. Chondrogenic derived-DSCMs remained compact and stronger during extraction and this made their handling easier. Ascorbic derived-DSCMs showed a different protein composition to chondrogenic-DSCMs. Bioactive characteristics analyzed were different depending on the cellular origin of DSCM and the method used to produce them. CONCLUSIONS: The DSCMs obtained in this work constitutes favorable structure- and growth factors providing a microenvironment which is very similar to that of native ECM, which results in enhanced biological potential of the MSCs and responsiveness to the induction of differentiation. We found differences between ascorbic derived-DSCMs and chondrogenic derived DSCMs. Our results suggest that the cell source used to produce DSCMs is highly related to the bioactive characteristics of DSCMs.