Polymer encapsulation of bacterial biosensors enables coculture with mammalian cells Moya-Ramírez, Ignacio Kotidis, Pavlos Marbiah, Masue Kim, Juhyun Kontoravdi, Cleo Polizzi, Karen Coculture Biosensor Bacteria Mammalian cells L-lactate Hydrogel encapsulation The authors gratefully acknowledge funding from the EPSRC Adventurous Manufacturing program (EP/T005297/1), the EPSRC Frontiers Engineering program (EP/K038648/1), and the BBSRC (BB/S006206/1). I.M.-R. also acknowledges funding from the Spanish Ministry of Science and Innovation through the project IJC2019-041817-I/AIE/10.13039/ 501100011033. Coexistence of different populations of cells and isolation of tasks can provide enhanced robustness and adaptability or impart new functionalities to a culture. However, generating stable cocultures involving cells with vastly different growth rates can be challenging. To address this, we developed living analytics in a multilayer polymer shell (LAMPS), an encapsulation method that facilitates the coculture of mammalian and bacterial cells. We leverage LAMPS to preprogram a separation of tasks within the coculture: growth and therapeutic protein production by the mammalian cells and L -lactate biosensing by Escherichia coli encapsulated within LAMPS. LAMPS enable the formation of a synthetic bacterial−mammalian cell interaction that enables a living biosensor to be integrated into a biomanufacturing process. Our work serves as a proof-of-concept for further applications in bioprocessing since LAMPS combine the simplicity and flexibility of a bacterial biosensor with a viable method to prevent runaway growth that would disturb mammalian cell physiology. 2024-09-24T07:55:49Z 2024-09-24T07:55:49Z 2022 journal article Moya-Ramírez, Ignacio et al. Polymer encapsulation of bacterial biosensors enables coculture with mammalian cells. ACS Synthetic Biology, 11, 3, 1303–1312. doi:10.1021/acssynbio.1c00577 https://hdl.handle.net/10481/94954 10.1021/acssynbio.1c00577 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Attribution-NonCommercial-NoDerivatives 4.0 Internacional ACS