Close to Real: Large‐Volume 3D Cell Spheroids on a Superamphiphobic Surface

Abstract

3D cell spheroids with native cell‐to‐cell junctions and cell‐extracellular matrix interactions can recapitulate the comprehensive physiological environments in vivo, providing new platforms for preclinical drug screening. However, the applications of 3D cell spheroids are limited by the unmatched size, deficient cell abundance, and short service time. In this work, a novel upward culture method on a porous silica superamphiphobic biointerface (SABI) is developed to fully overcome these technical hurdles. Cell suspensions maintain in a state of aquatic marble, accelerating cellular self‐aggregation. Contributing to the upward culture model, this strategy overcomes the shortages of size limitation and media refreshing in the classical methods. Herein, large‐volume 3D cell spheroids with the size of over 1500 µm can be efficiently established within 2 days and subsequently long‐term cultured for more than 20 days. For a close to real biomimetic microenvironment, breast tumor 3D spheroid is applied as a proof of concept, simultaneously employing MCF‐7 cell, cancer‐associated fibroblast (clone 8), and vascular endothelial cell (HUVEC). The large‐volume tumor spheroid with close‐to‐real microenvironment services as a unique paradigm for all‐in‐one biological relevant researches, including the capacity of in situ observation, long‐term incubation, drug screening, and recyclability.

Culture of cancer spheroids and evaluation of anti-cancer drugs in 3D-printed miniaturized continuous stirred tank reactors (mCSTRs)

“…Finally, at day 19, the outlet glucose concentration began to increase in a practically linear fashion to reach the values of the inlet condition at day 24 (1.06 ± 0.03 mg ml −1 ). This suggests a sharp decay in the metabolic activity of the spheroids under culture during this period (days [19][20][21][22][23][24]. As shown previously, this observation is consistent with the contraction in the volume and the decrease in viability of the spheroids examined at day 20.…”

“…Some novel medium-to high-throughput fabrication techniques are capable of producing homogeneous populations of cancer spheroids in the size range of 200-400 µm [18][19][20][21][22]. However, these small spheroids tend not to be physiologically relevant, as they do not fully represent the sizes of even the smallest clinically relevant avascular tumors found in patients, which normally exceed a size threshold of 400 µm [23][24][25]. This size differential is important because spheroids larger than 400-500 µm exhibit tumor-like oxygen gradients and hypoxic conditions in their cores [21,26], while hypoxia has been described as a contributor to the resistance to anti-cancer drugs exhibited by real tumors [21,27,28].…”

Pharmaceutical Approach to Develop Novel Photosensitizer Nanoformulation: An Example of Design and Characterization Rationale of Chlorophyll α Derivative

“…Regarding the selective screening of drug cytotoxicity, 3D spheroids are becoming more and more widespread compared to 2D models. Three-dimensional spheroids reflect all kinds of intercellular interactions, as well as the interaction of cells with the extracellular matrix providing more adequate modeling of specific tissue architecture [ 73 ].…”

Multidimensional controllable fabrication of tumor spheroids based on a microfluidic device

“…Then, the interactions between E-cadherin on adjacent cells promote compact spheroid formation. 16 Hanging droplets, 17,18 spinning flasks, 19 non-adherent substrates, 20,21 and force-driven methods 22,23 have been developed for mass manufacture and investigation of tumor spheroids. These methods force cells to aggregate by creating an inappropriate external adhesion environment, but cell-cell and cell-external interactions remarkably influence the generation of cell spheroids.…”