Expression pattern of histone H3 subtypes in articular chondrocytes

Three-dimensional cell culture used for tissue engineering has its own rules and directions. There is a deficiency of proliferative markers suitable for tissue engineering research when cells are cross-linked in network of fibers or suspended in hydrogel. It limits cell harvesting or impairs the flow to cell area conventional chemicals used as proliferative markers. According to current published data, the expression of replication-dependent histone H3 genes could be novel proliferative marker of cells. The intensive synthesis of H3 histones is tightly correlated with DNA synthesis and H3 mRNA is rapidly degraded when the S phase is completed or inhibited by cell cycle inhibitors. Based on this relation, non-dividing cells contain no H3 mRNA. The aim of the study was to determine expression pattern of replication-dependent H3 subtypes and tissue-specific H3/t subtype in normal human connective tissue cells. Analyzed cellular model was chondrocytes cell line due to the phenomenon that articular cartilage doesn't have natural ability to heal its injuries, consequently development of cartilage engineering is necessary. Evaluation of expression pattern was performed using Reverse Transcription PCR and reaction products were visualized on the gel electrophoresis. This study demonstrated that RT-PCR technique can be successfully used to study the expression of different histone H3 subtypes. Presented electrophoregram showed differential expression of the analyzed subtypes (no expression of H3/g and H3/t subtypes). Incubation with sodium butyrate and quantitative Real Time PCR enabled quantification of mRNA level of selected H3/d subtype. This part of study showed a significant reduction in the mRNA level of H3/d when the sodium butyrate was added. Obtained results indicated the possibility of using the expression of individual histone H3 subtype as a new proliferative marker.