Establishment of a Biophysical Effect-Based Platform for Rapid Expansion of Functional Mesenchymal Stem Cells

More than three hundreds allogenic mesenchymal stem/stromal cell (MSC) transplantations are ongoing in the clinical trials, and it is predictable that MSC transplantation can be a standard therapy in the near future. Due to a demand of large scaled cell number for clinical use, automatic cell culture may avoid the man made mistake and reduce the production cost. Currently, there is no commercialized automatic cell culture machine specific for MSC manufacturing. We have established a microfluidic MSC culture system and automatic cell loading by a robotic arm in the lab. The limitation for current machine is that contact-induced pre-mature senescence in MSCs is inevitable when confluent culture occurs, and MSC number can’t be expanded rapidly without passage the cell. In addition to overcome the cell number need in an automatic culture system, enhancement of cell activity represents promotion of therapeutic efficacy. In the past years, we are devoted to reinforce the activities of MSCs through biophysical effects triggered by physical stimuli. Shear stress induces F-actin rearrangement, which accompanies dynamic regulation of MSC activities. Change of F-actin orientation alters self-renewal, differentiation or paracrine ability of MSCs depending on different shear patterns. Besides, green light photo-irradiation enhances the directional MSC migration via activation of intracellular signaling during MSC homing. However, to combine the above research achievements in one automatic culture system for producing large-scaled MSCs with high quality is a big challenge for the science and our technology. The aim of this study is to construct an automatic photo-microfluidic cell culture machine to manufacture next generation MSCs. In this 5-year study, we will create multiple patterned cell loading devices and test the automatic cell loading into the loading device in the first year. The photo device and microfluidic device providing shear stress will be designed and photo-microfluidic system will be set up and tested in the second year. In the third year, we will optimize the initial seeding pattern of MSCs in an automatic culture system to prevent the confluence culture-induced pre-mature senescence. In the fourth year, the optimized shear condition to enhance immunomodulation, differentiation and paracrine cell protection ability in MSCs will be determined, respectively. In the fifth year, we will compare the effect of photo-stimulated directional migration on MSCs before and after cryopreservation, so that the optimal photo condition can be determined. At the end pf the study, a prototype will be fabricated. This is a study with high originality summarizing the research achievement in my lab in the past 10 years. This is also an important translational and applied research in developing the second generation MSCs like a target drug. The research achievement from this study may increase the therapeutic efficacy of MSC transplantation as well as reduce the risk and cost from man-made procedure, so that a large number of patients can benefit from MSC transplantation.