Plug-and-Play Lymph Node-on-Chip: Secondary Tumor Modeling by the Combination of Cell Spheroid, Collagen Sponge and T-Cells

Abstract

Towards the improvement of the efficient study of drugs and contrast agents, the 3D microfluidic platforms are currently being actively developed for testing these substances and particles in vitro. Here, we have elaborated a microfluidic lymph node-on-chip (LNOC) as a tissue engineered model of a secondary tumor in lymph node (LN) formed due to the metastasis process. The developed chip has a collagen sponge with a 3D spheroid of 4T1 cells located inside, simulating secondary tumor in the lymphoid tissue. This collagen sponge has a morphology and porosity comparable to that of a native human LN. To demonstrate the suitability of the obtained chip for pharmacological applications, we used it to evaluate the effect of contrast agent/drug carrier size, on the penetration and accumulation of particles in 3D spheroids modeling secondary tumor. For this, the 0.3, 0.5 and 4 μm bovine serum albumin (BSA)/tannic acid (TA) capsules were mixed with lymphocytes and pumped through the developed chip. The capsule penetration was examined by scanning with fluorescence microscopy followed by quantitative image analysis. The results show that capsules with a size of 0.3 μm passed more easily to the tumor spheroid and penetrated inside. We hope that the device will represent a reliable alternative to in vivo early secondary tumor models and decrease the amount of in vivo experiments in the frame of preclinical study.

Strategies for Anisotropic Fibrillar Hydrogels: Design, Cell Alignment, and Applications in Tissue Engineering

“…(h) Fluorescence images of chip with 4T1 cell tumor spheroid (Hoechst, blue fluorescence), T lymphocyte cells (Calcein Am, green fluorescence), and 4 μm polymer capsules (Cy5, red fluorescence). Reprinted with permission under a Creative Commons CC BY 4.0 license from ref . Copyright 2023 MDPI.…”

“…In a recent study, German et al. demonstrated a novel approach utilizing one-way freezing to create a collagen gel through the freezing of a reactive mixture . A mold consisting of an aluminum substrate and 3D printed walls from polylactide (PLA) was used to form an anisotropic collagen foam.…”

“…In a recent study, German et al demonstrated a novel approach utilizing one-way freezing to create a collagen gel through the freezing of a reactive mixture. 176 A mold consisting of an aluminum substrate and 3D printed walls from polylactide (PLA) was used to form an anisotropic collagen foam. Two wt % collagen gel in 0.02 M acetic acid was placed in the mold, where the directional crystallization was provided by the cooling of metal substrate with liquid nitrogen.…”

Mimicking blood and lymphatic vasculatures using microfluidic systems

“…Most models look at cancer and immune cell integration into the LN. 101–103 LECs tend to differentially express markers throughout the LN which could make modeling the endothelium a challenge. 104 For example, caveolin 1 appears in ceiling LECs in the subcapsular sinus, but tumor necrosis factor receptor superfamily member 9 (TNFRSF9) was seen in the floor LECs.…”

Evaluation of polymeric submicron microcapsule accumulation in cellular and intercellular space of 3D spheroids

“…Однако исследования, производимые большинством групп, направленные на изучение интернализации полимерных микрокапсул в клетки, связаны с классическими 2D клеточными культурами, которые не позволяют объективно оценивать эффективность используемых капсул в трехмерных биологических матрицах [11]. Одной из важных задач использования систем тестирования клеток in vitro является воссоздание микросреды in vivо [12]. Клетки в тканях окружены микросредой, состоящей из соседних клеток и внеклеточных матриц (ВМ), которые связанными с клеткам сигнальными путями, например, каскадом биохимических и механических сигналов.…”