Continuous human iPSC-macrophage mass production by suspension culture in stirred tank bioreactors
Macrophages derived from human induced pluripotent stem cells (iPSCs) have the potential to enable the development of cell-based therapies for numerous disease conditions. A protocol for the mass production of iPSC-derived macrophages (iPSC-Mac) in scalable suspension culture on an orbital shaker or in stirred-tank bioreactors (STBRs) have been developed by Dr. Lachmann and coworkers which have been recently published in Nature journal. This strategy is straightforward, robust and characterized by the differentiation of primed iPSC aggregates into ‘myeloid-cell-forming-complex’ intermediates by means of a minimal cytokine cocktail. In contrast to the ‘batch-like differentiation approaches’, myeloid-cell-forming-complex-intermediates are stably maintained in suspension culture and continuously generate functional and highly pure iPSC-Mac. Employing a culture volume of 120 ml in the STBR platform, ~1–4 × 107 iPSC-Mac can be harvested at weekly intervals for several months. The STBR technology allows for real-time monitoring of crucial process parameters such as biomass, pH, dissolved oxygen, and nutrition levels; the system also promotes systematic process development, optimization and linear upscaling. The process duration, from the expansion of iPSC until the first iPSC-Mac harvest, is 28 d. Successful application of the protocol requires expertise in pluripotent stem cell culture, differentiation and analytical methods, such as flow cytometry.
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