ALTEN: A High‐Fidelity Primary Tissue‐Engineering Platform to Assess Cellular Responses Ex Vivo

Abstract

To fully investigate cellular responses to stimuli and perturbations within tissues, it is essential to replicate the complex molecular interactions within the local microenvironment of cellular niches. Here, the authors introduce Alginate‐based tissue engineering (ALTEN), a biomimetic tissue platform that allows ex vivo analysis of explanted tissue biopsies. This method preserves the original characteristics of the source tissue's cellular milieu, allowing multiple and diverse cell types to be maintained over an extended period of time. As a result, ALTEN enables rapid and faithful characterization of perturbations across specific cell types within a tissue. Importantly, using single‐cell genomics, this approach provides integrated cellular responses at the resolution of individual cells. ALTEN is a powerful tool for the analysis of cellular responses upon exposure to cytotoxic agents and immunomodulators. Additionally, ALTEN's scalability using automated microfluidic devices for tissue encapsulation and subsequent transport, to enable centralized high‐throughput analysis of samples gathered by large‐scale multicenter studies, is shown.

GITR and TIGIT immunotherapy provokes divergent multi-cellular responses in the tumor microenvironment of gastrointestinal cancers

“…This feature enables culture in media free from cytokines such as IL-2 that are routinely used in maintaining T cells in culture for longer duration. As we have demonstrated previously, IL-2 can reprogram transcriptional T cell states (61). Our approach allows the evaluation of cell states in the original TME.…”

Performance comparison of high throughput single-cell RNA-Seq platforms in complex tissues

“…Complementary strategies, such as performing multi-omic studies where two or more molecular layers of information are investigated per cell 55 , or spatial transcriptomics 56 , as well as alternative methods for tissue preservation such as the ALTEN system 11 or parafilm fixation in combination with fixed RNA profiling 51,57 , may also help dissect the true heterogeneity of complex tissues and overcome the current limitations of single-cell RNAseq.…”

“…Single-cell transcriptomics provides an unprecedented resolution of the composition and functionality of cellular niches in homeostatic tissues and during disease 2,3 . For example, scRNAseq has been used to unravel multicellular dynamic processes during embryogenesis and cell differentiation, tissue regeneration and morphogenesis, disease initiation and progression, and response to stimuli including drug treatments [4][5][6][7][8][9][10][11][12] .…”

A review on biomimetic models based on microfluidic devices for drug research

“…PMMA (poly(methyl methacrylate)) and polycarbonate materials were each used in 2 papers [6,7,9,14], and following them, various other materials such as PEEK (polyetheretherketone) [15], UV resin [15], PET (polyethylene terephthalate) [9], GelMa (gelatin methacryloyl) [16], ECM gel [5], agarose [17] and polystyrene [18] were used in one paper each. Additionally, there were 2 papers in which the device material information was not specified [19,20]. Among the 19 papers, 5 described using devices made from two or more composite materials [5][6][7]9,15].…”

“…Only 1 paper was tributing lung cancer [20]. There was 1 paper targeting various types of cancers [19]. Of the 6 brain-related studies, 5 commonly addressed the blood-brain barrier (BBB) [7,8,10,15,37], while the remaining 1 study focused on the spheroid model [11].…”