Direct and Label-Free Cell Status Monitoring of Spheroids and Microcarriers Using Microfluidic Impedance Cytometry.

Lingyan Gong, Chayakorn Petchakup, Pujiang Shi, Pei Leng Tan, Lay Poh Tan, Chor Yong Tay, Han Wei Hou

Journal: Small (Weinheim an der Bergstrasse, Germany) 2021;17(21):e2007500

PMID: 33759381

Abstract

3D cellular spheroids/microcarriers (100 µm-1 mm) are widely used in biomanufacturing, and non-invasive biosensors are useful to monitor cell quality in bioprocesses. In this work, a novel microfluidic approach for label-free and continuous-flow monitoring of single spheroid/microcarrier (hydrogel and Cytodex) based on electrical impedance spectroscopy using co-planar Field's metal electrodes is reported. Through numerical simulation and experimental validation, two unique impedance signatures (|Z | (60 kHz), |Z | (1 MHz)) which are optimal for spheroid growth and viability monitoring are identified. Using a closed-loop recirculation system, it is demonstrated that |Z | increases with breast cancer (MCF-7) spheroid biomass, while higher opacity (impedance ratio |Z |/|Z |) indicates cell death due to compromised cell membrane. Anti-cancer drug (paclitaxel)-treated spheroids also exhibit lower |Z | with increased cell dissociation. Interestingly, impedance characterization of adipose-derived mesenchymal stem cell differentiation on Cytodex microcarriers reveals that adipogenic cells (higher intracellular lipid content) exhibit higher impedance than osteogenic cells (more conductive due to calcium ions) for both microcarriers and single cell level. Taken together, the developed platform offers great versatility for multi-parametric analysis of spheroids/microcarriers at high throughput (≈1 particle/s), and can be readily integrated into bioreactors for long-term and remote monitoring of biomass and cell quality.

Address: School of Mechanical and Aerospace Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798.; School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798.; School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798.; School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551.; Environmental Chemistry and Materials Centre, Nanyang Environment and Water Research Institute, 1 CleanTech Loop, CleanTech One, Singapore, 637141.; Energy Research Institute, Nanyang Technological University Singapore, 50 Nanyang Drive, Singapore, 637553.; School of Mechanical and Aerospace Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore, 639798.; Lee Kong Chian School of Medicine, Nanyang Technological University, 11 Mandalay Road, Singapore, 308232.; Critical Analytics for Manufacturing of Personalized Medicine, Singapore-Massachusetts Institute of Technology Alliance for Research and Technology, 1 CREATE Way, #10-01, CREATE Tower, Singapore, 138602.

DOI: 10.1002/smll.202007500