Comparison of extraction methods and culture medium for umbilical cord lining- and wharton's jelly-derived mesenchymal stromal cells

ConclusionSix donated UC samples were disinfected, cut into discs and frozen in cryopreservant containing 10% DMSO. Samples were thawed at 37°C, then incubated in either media for 3 - 4 days at 37°C in 5% CO2.MSC were extracted from the incubated tissues and differentially treated to generate every permutation of cell source, extraction method and culture media (total conditions: 8). At every passage, cell doubling rate was calculated and morphology was captured. At Passage 6 (P6), trilineage differentiation and karyotype were evaluated by StemPro® differentiation kits and G-banding.MSC extracted from all conditions displayed fibroblastic, spindle-shaped morphology and plastic-adherence. Only P6 MSC cultured in DMEM:F12 but not PPT-6 displayed enlarged and polygonal morphology. Overall doubling rate of PTT-6 cultured MSC was significantly faster than its DMEM:F12 counterpart (50.1 ± 9.7 h versus 98.4 ± 37.5 h, P < 0.05). Cell doubling rate was independent of either extraction method or tissue source.Tissue source and extraction method did not influence trilineage differentiation. PTT-6 favoured adipogenesis while MSC from DMEM:F12 were inclined towards osteogenesis. Both media allowed for chondrogenesis. No gross chromosomal abnormalities were observed after at least 10 population doublings.The patented PTT-6 medium maintained MSC stemness and significantly hastened UC-MSC expansion without detriment to trilineage differentiation or chromosomal stability. Further wor...

Source: Cytotherapy - May 24, 2019 Category: Cytology Source Type: research