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Bionanofabrication and Bionano Devices in Tissue Engineering and Cell Transplantation
Published in Anil Kumar Anal, Bionanotechnology, 2018
To cultivate the cell culture, primary cells, cells isolated directly from a body by biopsy, surgery, or autopsy, are cultured for a finite time to produce primary cell cultures, which might be slightly different from primary cells. Primary cell cultures are obtained via explant culture, which involves the removal of ECM by mechanical means such as mincing or by chemical means such as enzyme digestion followed by the incubation of tissue on growth surface along with growth medium. Under light microscopic examination, if cells are observed in an area surrounding tissue mounds, mounds are removed and remaining cells are allowed to proliferate and the cell culture obtained is known as primary cell cultures (Godbey 2014).
Determination of antibiotic impurities in good manufacturing practices-grade cell therapy medicinal products
Published in Preparative Biochemistry & Biotechnology, 2020
Olga Nehir Oztel, Seval Korkmaz, Erdal Karaoz
All samples of human Umbilical Cord Derived MSCs (UC-MSCs) as cell therapy medicinal products were manufactured at the cGMP-certified facility at Liv Hospital Center for Regenerative Medicine and Stem Cell Manufacturing (LivMedCell). Umbilical cords were obtained from consenting patients delivering full-term infants by Cesarean section, who faced no complications throughout pregnancy. Briefly, cords were rinsed using PBS (Invitrogen/Gibco, Paisley, UK), umbilical cord ven and arteries were then removed and the remaining tissue was minced into 1–2 mm3 mm pieces for explant culture. Culture was maintained in MSC NutriStem® XF Basal Medium and MSC NutriStem® XF Supplement Mix (Biological Industries, Cromwell, CT, USA) cell culture media supplied with 2% human serum (SeraCare, Milford, MA, USA) and 50 U/mL P/S (Biological Industries). Cells were grown in a humidified atmosphere containing 5% CO2 at 37 °C and were subcultured until the third passage. Culture medium is replaced every 5 d until fibroblast-like adherent cells reach 70% confluence. For MSC characterization and qualification, the cells were harvested at their third passages, and quality control (QC) tests were performed. In order to manufacture the cell based therapy products, flow cytometry analysis, cell viability assessment, mycoplasma detection, gene expression, telomerase activity, endotoxin and sterility tests were performed as “Valid Quality Control Tests”. Culture media were collected for the detection of antibiotic residues. All samples were tested under seven different groups as listed in Table 1.