The cells were subjected to induced differentiation using commercial kits (StemPro A10070-01, A10071-01, and A10072-01; Gibco) according to the manufacturer's instructions. Briefly, for the chondrogenic cell differentiation, the eBMmsc, eADmsc, and eUCmsc were plated in 5 μl droplets at a concentration of 1.6 × 107 cells/ml and cultured under standard conditions for 2 h; then, the differentiation medium was added. The medium was replaced every 4 days for 2 weeks. For the osteogenic differentiation, 5 × 103 cells/cm2 were plated and incubated for 24 h; then, the IMDM medium was replaced by a specific differentiation medium, which was replaced every 4 days for 2 weeks. For the adipogenic differentiation, 1 × 104 cells/cm2 were plated and incubated for 24 h; then, the IMDM medium was replaced by a specific differentiation medium, which was replaced every four days for two weeks. The cells in the control group were plated under the same conditions but were maintained in IMDM medium. At the end of the two-week period, the cells were fixed and stained using Sudan black dye (Sigma-Aldrich) for the adipogenesis differentiation detection, Alizarin red (Sigma-Aldrich) for the osteogenesis detection, and Alcian blue (Sigma-Aldrich) for the chondrogenesis differentiation detection.
Stempro a10070 01
Manufactured by Thermo Fisher Scientific
Sourced in United States
The StemPro A10070-01 is a laboratory instrument designed for cell culture applications. It is capable of performing automated cell culture tasks, such as cell seeding, media exchange, and cell harvesting. The device is intended to provide a controlled and consistent environment for cell growth and maintenance.
Automatically generated - may contain errors
2 protocols using stempro a10070 01
1
Differentiating Mesenchymal Stem Cells
2
Characterization and Differentiation of ADMSCs
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ADMSCs were obtained from the School of Regenerative Medicine, Bengaluru (India). Cells were isolated from the subcutaneous depots of healthy volunteers undergoing cosmetic surgeries. Passage 4 cells were characterized for the expression of CD90, CD73, CD105 (1:1000 dilution, PE Tagged), CD45, CD34, and HLA-DR (1:1000 dilution, FITC Tagged) (BD Biosciences, CA, USA) as described in our previous study.10 (link) Cells were further characterized for their chondrogenic (Stempro-A1007101, Gibco, MD, USA), adipogenic (Stempro-A1007001, Gibco, MD, USA), and osteogenic (Stempro-A1007201, Gibco, MD, USA) potential as shown earlier.22 (link)
The conditioned medium was prepared from ADMSCs as described earlier.10 (link) ADMSCs were seeded in 35 mm cell culture plates at the density of 1 × 106 cells per well. Once cells were 80%–85% confluent, fresh α-MEM medium (32561037, Gibco, MD, USA) (without serum) was added and incubated for 48 h at 37 °C & 5% CO2. After incubation, the culture medium was collected and filtered through 0.22 μmol/L filters. ADMSC-CM was stored at −80 °C until further use. It was diluted to 50% concentration with α-MEM (without serum) and used for all the experiments.
The conditioned medium was prepared from ADMSCs as described earlier.10 (link) ADMSCs were seeded in 35 mm cell culture plates at the density of 1 × 106 cells per well. Once cells were 80%–85% confluent, fresh α-MEM medium (32561037, Gibco, MD, USA) (without serum) was added and incubated for 48 h at 37 °C & 5% CO2. After incubation, the culture medium was collected and filtered through 0.22 μmol/L filters. ADMSC-CM was stored at −80 °C until further use. It was diluted to 50% concentration with α-MEM (without serum) and used for all the experiments.
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