Production of ADSC-CM
MYCOPLASMA, STERILITY, -20
ADSC-CM was produced by the Stem Cell Medical Technology Unit of Dr. Cipto Mangunkusumo Hospital. ADSC-CM used in this study was allogeneic, formed by the culture of human mesenchymal stem cells (MSC). These mesenchymal stem cells originated from adipose tissue which has been screened previously for infectious pathogens. After successful isolation of MSCs, these stem cells were multiplied in a normoxic condition until it reached the fifth passage, with a culture confluence of at least 80%. The conditioned media were then obtained by removing the stem cells.Following a round of centrifugation to remove existing debris, ADSC-CM was stored frozen at -20 °C and slowly thawed at 4 °C overnight before undergoing filtration using a 0.22 µm syringe filter. Further processing for concentrated ADSC-CM was conducted using the Spin-X UF 500 Concentrator (Corning). Both non-concentrated and concentrated ADSC-CM was subjected to sterility tests to exclude the presence of bacteria, fungi, and mycoplasma endotoxins, as well as a potency test to evaluate the number of proteins and growth factors.