Life Span of the Chondrocytes from Human Umbilical Cord Derived-Mesenchymal Stem Cell Differentiation

Abstract

Problem in growth might need to be sought deeper inside the cells to enhance better management. There was a lack of study done in chondrocytes to determine growth problem mechanism. We sought to see the chondrocyte’s life span by observing the cells characteristic in a small container over long period. Human mesenchymal stem cell-derived MSC underwent chondrocyte differentiation procedure in a 96 wells-plate using chondrocyte/osteocyte differentiation medium followed by basal medium. A morphological observation and MTT assay was done to observe apoptosis in chondrocyte in the different wells. MTT assay absorbance was used to determine number of apoptosis. Statistical analysis was done to assess the different of mean absorbance values on different days of observation using Anova folowed by post hoc analysis in SPSS IBM package version 24 , with significance determined at P<0.05. Chondrocytes was confirmed with 1% Alcian blue staining and showed characteristic larger then its parent MSC. The MTT assay showed that chondrocyte absorbance reduce from day 3 through day 14, which was different significantly (anova, P=0.03) and day 14 was the most differed compared to day 7 and day 10 (P=0.02, P=0.017 and P=0.013, respectively). The 2D monolayer chondrocyte underwent total apoptosis on day 14 which can set the limit for observation in further long-term observational cell study.