ADIPOSE-DERIVED MESENCHYMAL CELLS YIELD UNDER DIFFERENT CELL EXTRACTION PROTOCOLS

Abstract

Objective: To establish a protocol for extraction of adipose-derived mesenchymal cells, evaluating the cell yield in two methods of tissue enzymatic digestion. Material and Methods: Fragments of adipose tissue were removed from the inguinal region of mice and processed by two different protocols: Group 1 - enzymatic digestion with collagenase I; Group 2 - enzymatic digestion with collagenase I and trypsin/EDTA, both for 1 hour at 37°C. In the third passage (P3), cells from both groups were counted in a Neubauer chamber for evaluation of cell proliferation, and growth curves were plotted at intervals of 24, 48 and 72 hours. The DAPI nuclear staining was performed in both groups in a 72-hour interval to evaluate the presence of nuclear morphological damage. Data were analyzed statistically with a significance level of 5%. Results: The two groups showed an upward pattern of cell growth. The mean cell counts showed a higher cell proliferation in group I, with a statistically significant difference between groups in intervals of 48 and 72 hours (p<0.05). Nuclear morphological changes were not observed in the groups. Conclusion: The enzymatic digestion of adipose tissue by collagenase I, without association with trypsin, improves the yield of mesenchymal cells, what suggests the choice of that protocol in experiments with this cell type. DESCRIPTORS: Adipose Tissue. Cell Culture Techniques. Cell Proliferation.