The standard diabetes therapy with insulin is a burden for the patient. Alternative therapy forms such as the replenishment of destroyed beta cells with insulin-producing cells are on the rise. The main obstacles for such therapies are sources of sufficient insulin-producing cells as well as high amounts of high-quality cells to satisfy the needs for cell therapies with 106-1010 cells per dose. In standard monolayer/2D cultures, mature beta cells grow slowly and lose their functionality (e.g. no secretion of insulin anymore). Therefore, a more complex and supporting 3D environment, such as the cultivation as agglomerates/pseudoislets, can fasten beta cell growth and restore e.g. glucose-dependent insulin secretion. Nonetheless, mono-cultured beta cells are not very active in vitro. As in nature beta cells are surrounded by several other cells types, a co-cultivation of beta cells with e.g. mesenchymal stromal cells (hMSCs) can improve the pseudoislets’ survival and increase the beta cell functionality. The positive effect of hMSCs is driven by cell-cell interactions and secreted trophic factors. like VEGF or HGF. Here, we describe a bioreactor-based co-cultivation concept of hMSCs with insulin-producing beta cell lines. We investigated several co-cultivation types e.g. direct, indirect and mixed. The direct co-cultivation enables direct cell-cell interactions, whereas the indirect co-cultivation physically separates the cells and any effect is only given through secreted factors. We determined the effect of different cell ratios and a medium composition on the beta cells functionality. For the bioreactor setup, the cells were either expanded on microcarriers or in agglomerates. We integrated online monitoring tools to control cell growth within the bioreactor. We further want to investigate the effect of the bioreactor hydrodynamics on the cells. Our aim is the development of a high-mass co-cultivation process which satisfies the needs of both cell types and meets the requirements of good manufacturing practice and process analytical technology.

Co-cultivation: A powerful tool for insulin-producing beta cell manufacturing

2019 Spring National Meeting

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