Cell detachment rates and confluence of fibroblast and osteoblast cell culture using different washing solutions

Abstract

The advancements in the cell culture studies have led to the development of regenerative medicine concept. The aim of this study is to compare the effectiveness of some washing solutions, including phosphate buffered saline (PBS), sodium chloride (NaCl), and ringer's lactate (RL) on the rate of detachment and confluency in fibroblast and osteoblast cell culture. Baby Hamster Kidney 21 clone 13 (BHK21/C13) fibroblast cells and 7F2 osteoblast were cultured on T25 flasks for 3-4 days. Three treatment groups were classified on the basis of different washing solutions used in the moment before trypsinization: PBS, 0.9% NaCl, and RL. Each group was measured for the detachment rate and cell confluence. The measurement was done in 2 passage numbers. The use of PBS, NaCl, and RL washing solution showed that detachment time was less than 5 minutes for the fibroblasts and 3 minutes for the osteoblasts. There was a significant difference in the rate of fibroblast cell detachment (p=0.006) and osteoblast (p=0.016). The capability of fibroblasts and osteoblasts to achieve a confluence of 106 cells/well on the first and second measurements was almost the same between the washing solution groups. The use of physiological 0.9% NaCl solution as a washing solution in fibroblast and osteoblast cell culture has almost the same effectiveness as PBS to help accelerate cell detachment in less than 5 minutes without influencing the capability of cells to proliferate.

Effect of Silicone Rubber/Polyvinyl Alcohol /Zirconium Oxide Compound on Mesenchymal Stromal Cells

“…This concentration is reflective of the constituents released from the initial contact, as opposed to the direct method, where the maximal concentration is achieved only at the end of the treatment period, normally 24 h ( 14) . This standard recommends Phosphate Buffer saline (PBS) as a buffer solution which is isotonic and non-toxic and aims to maintain cell osmolarity and not interfere with tissue viability (25). MTT undergoes enzymatic reduction to purple formazan in metabolically active cells Assay is then done by comparing cell exposure to a substance and measuring the decrease in optical density.…”