In Vitro Study of a Novel -Based Collagenase for Future Medical Application.

Mesenchymal stem cells extracted from adipose tissue are particularly promising given the ease of harvest by standard liposuction and reduced donor site morbidity. This study proposes a novel enzymatic method for isolating stem cells using collagenase, obtaining a high-quality product in a reduced time. Initially, the enzyme concentration and incubation time were studied by comparing cellular yield, proliferation, and clonogenic capacities. The optimized protocol was phenotypically characterized, and its ability to differentiate in the mesodermal lineages was evaluated. Subsequently, that protocol was compared with two -based collagenases, and other tests for cellular integrity were performed to evaluate the enzyme's effect on expanded cells. The best results showed that using a concentration of 3.6 mg/mL collagenase allows extracting stem cells from adipose tissue after 20 min of enzymatic reaction like those obtained with -based collagenases after 45 min. Moreover, the extracted cells with collagenase presented the phenotypic characteristics of stem cells that remain after culture conditions. Finally, it was seen that collagenase does not reduce the vitality of expanded cells as -based collagenase does. These findings suggest that collagenase has great potential in regenerative medicine, given its degradation selectivity by protecting vital structures for tissue restructuration.