Human chondrocyte-conditioned medium promotes chondrogenesis of bone marrow stem cells
Article Category: Original article
Publicado en línea: 13 jul 2020
Páginas: 9 - 18
DOI: https://doi.org/10.1515/abm-2020-0003
Palabras clave
© 2020 Rabiatul Adawiyah Razali et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.
Background
Cell-based therapy for osteoarthritis requires culturing of good quality cells, especially with a chondrogenic lineage, for implantation.
Objective
To investigate the ability of chondrocyte-conditioned medium (CCM) to induced chondrogenesis.
Methods
Bone marrow mesenchymal stem cells (BMSCs) were subjected to chondrogenic induction using CCM and chondrocyte induction medium (CIM). The optimal condition for the collection of CCM was evaluated by quantifying the concentration of secreted proteins. The chondrogenic efficiency of BMSCs induced by CCM (iCCM) was evaluated using immunocytochemical analysis, Safranin-O staining, and gene expression.
Results
Protein quantification revealed that CCM obtained from cells at passage 3 at the 72 h collection point had the greatest amount of protein. Supplementation of CCM results in the aggregation of BMSCs; however, no clumping was visible as in iCIM. The expression of collagen type 2 was detected as early as day 7 for all groups except for non-induced BMSCs; however, the level of expression decreased with culture time. Similarly, all tested groups showed positive staining for Safranin-O as early as day 7. The induction of BMSCs by CCM caused the down-regulation of collagen type 1, along with the up-regulation of the collagen type 2, ACP and SOX9 genes.
Conclusion
The optimum CCM to induce BMSC into chondrocytes was collected at passage 3 after 72 h and was used in a 50:50 ratio of CCM to fresh medium.