Comparison of dissociation capability of papain (Carica papaya L.) and trypsin on rat bone marrow mesenchymal stem cells (rBMMSCs) culture (preliminary study)

Abstract

Trypsin is an important enzyme that used for cell culture dissociation, which is still dominant in stem cell application. On stem cell transplantation, some people disagree if its product applicated in the human body because trypsin is an animal product. Papain is an enzyme produced by Carica papaya and nontoxic in cell culture. Papain is successful to dissociate cortical – neuron cell culture. This research is preliminary research to using papain in dissociation on rat bone marrow mesenchymal stem cells (rBMMSCs). The aim of this study were to determine the dissociation capability of papain, find the optimal concentration of papain to dissociation rBMMSCs compared with trypsin and characterize rBMMSCs after dissociating with papain. rBMMSCs was isolated from rat bone marrow and after the 1st passage of rBMMSCs were seeding 50.000 cells/mL in each well in 24-well plate. The cells cultured for 5 days and dissociated with trypsin and papain (20 µg/mL, 40 µg/mL, 60 µg/mL and 80 µg/mL). Characterization of rBMMSCs after dissociation was done by observed cells number and flow cytometry analysis with markers CD90 and CD34. Statistical analysis using the SPSS program with one-way ANOVA test. The results showed that papain able to dissociate culture cells become a single cell as good as trypsin. The optimum concentration of papain is 80 µg/mL for 5 minutes to make perfectly dissociated rBMMSCs.