Gravity-Based Flow Efficient Perfusion Culture System for Spheroids Mimicking Liver Inflammation.

Young-Su Kim,Jae-Wook Lee,Abdul Rahim Chethikkattuveli Salih,Arun Asif,Ki-Nam Hyun,Kyung-Hyun Choi

doi:10.3390/biomedicines9101369

Abstract

The spheroid culture system provides an efficient method to emulate organ-specific pathophysiology, overcoming the traditional two-dimensional (2D) cell culture limitations. The intervention of microfluidics in the spheroid culture platform has the potential to enhance the capacity of in vitro microphysiological tissues for disease modeling. Conventionally, spheroid culture is carried out in static conditions, making the media nutrient-deficient around the spheroid periphery. The current approach tries to enhance the capacity of the spheroid culture platform by integrating the perfusion channel for dynamic culture conditions. A pro-inflammatory hepatic model was emulated using a coculture of HepG2 cell line, fibroblasts, and endothelial cells for validating the spheroid culture plate with a perfusable channel across the spheroid well. Enhanced proliferation and metabolic capacity of the microphysiological model were observed and further validated by metabolic assays. A comparative analysis of static and dynamic conditions validated the advantage of spheroid culture with dynamic media flow. Hepatic spheroids were found to have improved proliferation in dynamic flow conditions as compared to the static culture platform. The perfusable culture system for spheroids is more physiologically relevant as compared to the static spheroid culture system for disease and drug analysis.

Highlights

Spheroid culture systems provide the opportunity to formulate sophisticated threedimensional (3D) tissues to mimic the organ-specific microenvironment and pathophysiology
The organ-on-a-chip and spheroid culture systems are evolving to better mimic human pathophysiology, which is being enhanced by the integration of sensors and robotics [1,2,3,4,5,6,7,8]
Albumin staining was assessed owing to its essential role in indicating hepatocyte functionality

Summary

Introduction

Spheroid culture systems provide the opportunity to formulate sophisticated threedimensional (3D) tissues to mimic the organ-specific microenvironment and pathophysiology. The organ-on-a-chip and spheroid culture systems are evolving to better mimic human pathophysiology, which is being enhanced by the integration of sensors and robotics [1,2,3,4,5,6,7,8]. Among MPS, spheroids are emerging with the potential to mimic human tissues in a 3D shape and function [9,10,11]. Spheroids are generated using multiple methods including culture on ultra-low attachment (ULA) surfaces, bioreactors [12], and cell aggregation in hanging drops [13]. Creating a non-adherent surface averts the attachment of cells to the substrate derives spheroid development. Liquid overlay techniques have been used to establish a nonadherent surface for cells to form 3D spheres, which include poly-2-hydroxyethyl methacrylate (poly-HEMA), pluronic acid, or 1–2% agarose coating on the substrate surface [16]

Methods

Results

Discussion

Conclusion