The study of anterior cruciate ligament fibroblasts and synovium-derived mesenchymal stromal cells direct co-culture system on chitosan substrate

In this study, we isolated cells from human anterior cruciate ligament (ACL) and synovial membrane. Further we demonstrate that these cells from synovial membrane are similar with mesenchymal stem cells (MSCs) by flow cytomerty analysis and differentiation assay. So we can call the cell is synovium-derived stromal cells (SDMSCs).

Chitosan is a natural, biodegradable, biocompatible, non-toxicity and the U.S. Food and Drug Administration (FDA) approval of polysaccharides. In this study, chitosan was used to as substrate. When anterior cruciate ligament fibroblasts (ACLFs) and SDMSCs were cultured on chitosan substrate, the cells morphology was suspension and accumulation. Then we culture ACLFs and SDMSCs on chitosan substrate at direct co-culture system to explore the interaction between two cells, and TCPS as the control group.

Furthermore, we labeled the two cells by CellTrackerTM and used the inverted fluorescence microscope to observe the two cells pattern on substrate after direct co-cultured. Besides, we used time-lapse to observe the two cells pattern when the two cells direct co-culture on substrate dependent with time.

Finally, we used the flow sorter system to separate the two cells on the substrate after direct co-culture. After RNA was extracted, we use real-time quantitative polymerase chain reaction (RT-PCR) to observe genes expression of ACLFs and SDMSCs on different substrates after direct co-culture.