Optimization of culture of mesenchymal stem cells: a comparison of conventional plate and microcarrier cultures

Objectives There has been increasing interest in mesenchymal stem cells ( MSC s) because of their potential use for regenerative therapy; however, there is no well‐defined protocol for MSC s culture. This study compares techniques of conventional plate and microcarrier culturing of MSC s. Methods and results Here, different conditions for isolation and expansion of rat MSC s have been examined and it was found that plating density and plating time in primary culture played important roles for culture of these rat MSC s. When plated at 10 8 /cm 2 density for 72 h, in primary culture, recycling stem cells ( RS cells) predominated, and characteristics of rat MSC s (including morphology, growth rate, phenotype and differentiation potentials) remained stable during expansion until passage 14. For subculture of the cells, it was found that their growth rate when incubated at 33 °C was higher than those incubated at 37 °C, and maximal increase was 10‐ and 6‐fold respectively. When cultured using microcarriers, at a density of 1 × 10 5 /mg beads, growth kinetics, phenotype and differentiation potentials also remained constant for cells between passage 2nd and 14th; their maximal number increased 16‐fold. Conclusions Compared to conventional plate culture, culture using gelatine porous microcarrier C ultispher‐S was superior for large‐scale production of rat MSC s.