Development of a cell therapy set prototype for autologous iPS cells.

Background: Cell therapy using differentiated cells with less immune rejection of the patient's own cells can be realized by generating therapeutic cells from induced pluripotent stem cells (iPSCs). Current methods to produce iPSCs and differentiated cells in cell culture and processing facilities under GMP control typically involve 5-6 people working in one room. This process is suitable for large-scale production, but not for custom manufacturing of autologous iPSCs. A cell manufacturing process suitable for the production of autologous iPSCs can be implemented by simplifying aseptic operations, using small cell culture bags to enable small batch production, and providing a cell processing platform that integrates successive cell processing steps by building a connected loop of small cell culture bags.

Objective: We are developing a closed culture system to produce therapeutic cells using iPSCs derived from the patient's own cells. Cell Therapy Applications for Autologous iPSCs is a closed culture kit with 10 culture ports (small cell culture bags with many connecting tubes for cell processing) for each culture step.

Study design: Cells are moved from one culture port to the next at each step of the process. We initially designed our own culture port, which is a cell culture site in the cell therapy kit. Manufacturing was outsourced to CellBios (India). The ports were designed according to a series of 10 linked steps as follows: vector infection (2 hours), vector dilution, iPSC establishment (14 days), iPSC proliferation 1 (7 days), iPSC proliferation 2 (7 days), differentiation inducing solution A (0-10 days), differentiation inducing solution B (0-10 days), differentiation inducing solution C (0-10 days) and final formulation process of the cell product.

Results: During iPSC derivation and differentiation induction, cells are attached to atelocollagen beads so that the cells and beads can move between culture processes without cell detachment. We conducted a pilot study to determine the effect of the beads on iPSC differentiation. In the pilot study, we first established iPSCs on atelocollagen beads using Peripheral Blood Mononuclear Cells (PBMCs) and induced the differentiation of different cells (cardiomyocytes, pancreatic progenitor cells, and dopaminergic neurons). Marker expression of the differentiated cells was evaluated.

Conclusion: A series of processes were reproduced in a closed culture system to study cell characteristics.