The iPSC secretome is beneficial for in vitro propagation of primary osteoarthritic chondrocytes cell lines

• Published in: Biochemical and biophysical research communications 2024-10, Vol.730, p.150392, Article 150392
• Main Authors: Rosochowicz, Monika A., Lach, Michał S., Richter, Magdalena, Jagiełło, Inga, Suchorska, Wiktoria M., Trzeciak, Tomasz
• Format: Article
• Language: English
• Subjects: Cell Culture Techniques - methods; Cell Line; Cell Proliferation; Cells, Cultured; Cellular Senescence; chondrocytes; Chondrocytes - cytology; Chondrocytes - metabolism; Conditioned medium; Culture Media, Conditioned - metabolism; Culture Media, Conditioned - pharmacology; Humans; Induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Osteoarthritis; Osteoarthritis - metabolism; Osteoarthritis - pathology; Reactive Oxygen Species - metabolism; Secretome - metabolism
• ISSN: 0006-291X; 1090-2104; 1090-2104
• DOI: 10.1016/j.bbrc.2024.150392

One of the obstacles to autologous chondrocyte implantation (ACI) is obtaining a large quantity of chondrocytes without depletion of their properties. The conditioned medium (CM) from different subpopulations of stem cells (mesenchymal stromal cells (MSC) or induced pluripotent stem cells (iPSC)) could be a gamechanger. MSCs' potential is related to the donor's health and age, which could be omitted when, as a source, iPSCs are used. There is a lack of data regarding their use in the chondrocyte culture expansion. Thus, we wanted to verify whether iPSC-CM could be beneficial for the cell culture of primary chondrocyte cells. We added the iPSC-CMs from GPCCi001-A and ND 41658*H cells to the culture of primary chondrocyte cell lines isolated from OA patients (n = 6) for other two passages. The composition of the CM was evaluated using Luminex technology. Then, we analysed the senescence, proliferation rate and using flow cytometry: viability, distribution of cell cycle phases, production of reactive oxygen species (ROS) and double-strand breaks. The cartilage-related markers were evaluated using Western blot and immunofluorescence. Additionally, a three-dimensional cell culture was used to determine the potential to form cartilage particles. iPSC-CM increased proliferation and diminished cell ROS production and senescence. CM influenced the cartilage-related protein expression and promoted the growth of cartilage particles. The cell exposed to CM did not lose the ECM proteins, suggesting the chondroprotective effect for prolonged culture time. Our preliminary results suggest a beneficial effect on maintaining chondrocyte biology during in vitro expansion. [Display omitted] •iPSC secretome a new source for the management of degenerative diseases such as OA.•iPSC conditioned medium has a beneficial effect on the biology of chondrocytes.•CM increased proliferation, reduced ROS level and senescence of OA chondrocytes.•CM promoted the growth of cartilage particles in 3D culture.