Cell pausing method for adipose tissue derived mesenchymal stem cells: comparison of Petaka G3 and ordinary flask

Abstract

Background: Various methods of cell pausing were applied to prevent proliferation of cultured cells, which either required special medium, using complicated encapsulation method, or using a special flask (Petaka G3 flask). Therefore, this study aimed to develop an easy and economical method of cell pausing method for adipose tissue derived mesenchymal stem cells (AT-MSCs) using ordinary basal medium and flask that yielded AT-MSCs, which were compliant with the requirements of International society for cell therapy (ISCT). Methods: We assessed cell numbers and viabilities, and percentages of CD73, CD90, CD105, and lineage negative, as well as differentiation potentials of AT-MSCs, which were cultured until 80% confluent and then placed at 4 °C or room temperature for three to seven days. We assessed those parameters between fully filled and sealed ordinary flasks compared to Petaka G3 flasks. Results: Cell numbers, viabilities, and percentages of CD73, CD90, CD105, and lineage negative, as well as differentiation potentials of AT-MSCs at room temperature were better than at 4 °C, and at room temperature, results of ordinary flasks were comparable to Petaka G3 flasks. Conclusion: We have developed an easy and economical method of cell pausing method for adipose tissue derived mesenchymal stem cells (AT- MSCs), using ordinary basal medium and flask.