bims-orenst 2022-08-14 papers

Issue of 2022–08–14
four papers selected by

Front Bioeng Biotechnol. 2022 ;10 945149

A multi-organ-on-chip to recapitulate the infiltration and the cytotoxic activity of circulating NK cells in 3D matrix-based tumor model.

Monica Marzagalli, Giorgia Pelizzoni, Arianna Fedi, Chiara Vitale, Fabrizio Fontana, Silvia Bruno, Alessandro Poggi, Alessandra Dondero, Maurizio Aiello, Roberta Castriconi, Cristina Bottino, Silvia Scaglione.

The success of immunotherapeutic approaches strictly depends on the immune cells interaction with cancer cells. While conventional in vitro cell cultures under-represent the complexity and dynamic crosstalk of the tumor microenvironment, animal models do not allow deciphering the anti-tumor activity of the human immune system. Therefore, the development of reliable and predictive preclinical models has become crucial for the screening of immune-therapeutic approaches. We here present an organ-on-chip organ on chips (OOC)-based approach for recapitulating the immune cell Natural Killer (NK) migration under physiological fluid flow, infiltration within a 3D tumor matrix, and activation against neuroblastoma cancer cells in a humanized, fluid-dynamic environment. Circulating NK cells actively initiate a spontaneous "extravasation" process toward the physically separated tumor niche, retaining their ability to interact with matrix-embedded tumor cells, and to display a cytotoxic effect (tumor cell apoptosis). Since NK cells infiltration and phenotype is correlated with prognosis and response to immunotherapy, their phenotype is also investigated: most importantly, a clear decrease in CD16-positive NK cells within the migrated and infiltrated population is observed. The proposed immune-tumor OOC-based model represents a promising approach for faithfully recapitulating the human pathology and efficiently employing the immunotherapies testing, eventually in a personalized perspective. An immune-organ on chip to recapitulate the tumor-mediated infiltration of circulating immune cells within 3D tumor model.

Keywords: 3D human tumor model; cells infiltration; cells migration; immune-organ-on-chip; natural killer cells; neuroblastoma

DOI: https://doi.org/10.3389/fbioe.2022.945149

Anal Chim Acta. 2022 Aug 15. pii: S0003-2670(22)00664-X. [Epub ahead of print]1221 340093

A whole-thermoplastic microfluidic chip with integrated on-chip micropump, bioreactor and oxygenator for cell culture applications.

Amir Reza Ameri, Armin Imanparast, Mohammad Passandideh-Fard, Seyed Ali Mousavi Shaegh.

Microfluidics provides enabling platforms for various cell culture, drug testing and synthesis of drug carriers using chip-based microsystems. In this study, we present a novel integrated whole-thermoplastic microfluidic chip to provide a platform for on-chip cell culture at static and dynamic conditions. The whole chip was made of polymethyl methacrylate (PMMA) and thermoplastic polyurethane (TPU) using high precision micromilling and laser micromachining, assembled by thermal fusion bonding. Prior to fabricate the integrated microchip, a pneumatic solo diffuser-nozzle micropump was fabricated and characterized to evaluate its functionality for on-chip pumping. Then the micropump was integrated with a microbioreactor and an oxygenator in a microchip for flow pumping required for on-chip cell culture. Oxygenator, made of a thin TPU membrane and a reservoir, was implemented in the microchip because of low oxygen permeability of PMMA. To design the oxygenator for sufficient oxygen delivery to the chip, numerical simulation was performed using COMSOL Multiphysics® to evaluate oxygen concentration distribution inside the microchip. Finally, the diffuser-nozzle micropump was integrated with the oxygenator and a bioreactor on the microchip for cell culture with on-chip pumping. Culture of DFW cells was performed on the integrated chip for three days, and cell survival was evaluated with Trypan Blue assay. The findings reveal that the proposed integrated chip with on-chip pumping could be employed for conducting various cell culture studies.

Keywords: Diffuser-nozzle micropump; Microfluidics; Numerical simulation; On-chip cell culture; Thermoplastic

DOI: https://doi.org/10.1016/j.aca.2022.340093

Front Bioeng Biotechnol. 2022 ;10 908848

A three-dimensional culture system for generating cardiac spheroids composed of cardiomyocytes, endothelial cells, smooth-muscle cells, and cardiac fibroblasts derived from human induced-pluripotent stem cells.

Asher Kahn-Krell, Danielle Pretorius, Bijay Guragain, Xi Lou, Yuhua Wei, Jianhua Zhang, Aijun Qiao, Yuji Nakada, Timothy J Kamp, Lei Ye, Jianyi Zhang.

Cardiomyocytes (CMs), endothelial cells (ECs), smooth-muscle cells (SMCs), and cardiac fibroblasts (CFs) differentiated from human induced-pluripotent stem cells (hiPSCs) are the fundamental components of cell-based regenerative myocardial therapy and can be used as in-vitro models for mechanistic studies and drug testing. However, newly differentiated hiPSC-CMs tend to more closely resemble fetal CMs than the mature CMs of adult hearts, and current techniques for improving CM maturation can be both complex and labor-intensive. Thus, the production of CMs for commercial and industrial applications will require more elementary methods for promoting CM maturity. CMs tend to develop a more mature phenotype when cultured as spheroids in a three-dimensional (3D) environment, rather than as two-dimensional monolayers, and the activity of ECs, SMCs, and CFs promote both CM maturation and electrical activity. Here, we introduce a simple and reproducible 3D-culture-based process for generating spheroids containing all four cardiac-cell types (i.e., cardiac spheroids) that is compatible with a wide range of applications and research equipment. Subsequent experiments demonstrated that the inclusion of vascular cells and CFs was associated with an increase in spheroid size, a decline in apoptosis, an improvement in sarcomere maturation and a change in CM bioenergetics.

Keywords: biomanufacturing; cardiomyocyte; maturation; organoids; pluripotent stem cell; suspension culture

DOI: https://doi.org/10.3389/fbioe.2022.908848

Clin Transl Sci. 2022 Aug 12.

A proof of concept using the Ussing chamber methodology to study pediatric intestinal drug transport and age-dependent differences in absorption.

Eva J Streekstra, Márton Kiss, Jeroen van den Heuvel, Johan Nicolaï, Petra van den Broek, Sanne M B I Botden, Martijn W J Stommel, Lara van Rijssel, Anna-Lena Ungell, Evita van de Steeg, Frans G M Russel, Saskia N de Wildt.

Little is known about the impact of age on the processes governing human intestinal drug absorption. The Ussing chamber is a system to study drug transport across tissue barriers, but it has not been used to study drug absorption processes in children. This study aimed to explore the feasibility of the Ussing chamber methodology to assess pediatric intestinal drug absorption. Furthermore, differences between intestinal drug transport processes of children and adults were explored as well as the possible impact of age. Fresh terminal ileal leftover tissues from both children and adults were collected during surgery and prepared for Ussing chamber experiments. Paracellular (enalaprilat), transcellular (propranolol), and carrier-mediated drug transport by MDR1 (talinolol) and BCRP (rosuvastatin) were determined with the Ussing chamber methodology. We calculated apparent permeability coefficients and efflux ratios and explored their relationship with postnatal age. The success rate for the Ussing chamber experiments, as determined by electrophysiological measurements, was similar between children (58%, N = 15, median age: 44 weeks; range 8 weeks to 17 years) and adults (67%, N = 13). Mean serosal to mucosal transport of talinolol by MDR1 and rosuvastatin by BCRP was higher in adult than in pediatric tissues (p = 0.0005 and p = 0.0091). In contrast, within our pediatric cohort, there was no clear correlation for efflux transport across different ages. In conclusion, the Ussing chamber is a suitable model to explore pediatric intestinal drug absorption and can be used to further elucidate ontogeny of individual intestinal pharmacokinetic processes like drug metabolism and transport.

DOI: https://doi.org/10.1111/cts.13368