Effect of human articular chondroprogenitor derived conditioned media on chondrogenic potential of bone marrow derived mesenchymal stromal cells

Abstract

Background: Current research in cartilage repair is focusing on the use of soluble factors released by cells during expansion, either as stand-alone therapy or as conditioned media (CM) supplements to optimize cellular phenotype for in-vivo transplantation. The present study aimed at utilizing spent media from cultured human articular cartilage derived chondroprogenitors and assessing their influence on bone marrow mesenchymal stem cell (BM-MSC) growth and phenotype. Methodology: CM was collected from passage 2 chondroprogenitors and evaluated for latent TGFβ1 levels. Passage 3 BM-MSCs were divided into two groups and cultured with either a) standard expansion medium (EM-group) or b) EM supplemented with 50% CM (CCM-group). At sub-confluence both groups were assessed for population doubling, migration assay, cell surface markers, gene expression for chondrogenic and hypertrophy markers. Additionally, pellet cultures were subjected to chondrogenic differentiation and analyzed by Alcian blue stain. Results: On analysis of proliferation and migration, CCM-group showed comparable results to EM in terms of population doubling and cell movement toward scratched area. Similarly, use of spent medium did not affect the surface protein or gene expression profile of BM-MSCs with similar flow cytometric and mRNA results seen in both groups. Glycosaminoglycan deposition (Alcian blue) was seen in the CCM-group, comparable to the EM-group. Conclusion: This preliminary report provided valuable information on the influence of unfractionated CCM on BM-MSC characteristics which may be further optimized for cartilage regeneration by the use of purified components such as exosomes, micro-vesicles, and concentrated trophic factors in future.