Preparation of liver spheroids incorporated with PFC- MPs
Creation of spheroids and incorporation with MP followed procedures
recently published.[16] Briefly, HepG2 and HSC
cells were used for in vitro experimentation. Low glucose
Dulbecco’s Modified Eagle’s Medium with L-glutamine and sodium
bicarbonate (DMEM; Sigma Aldrich, cat#D6046) supplemented with fetal
bovine serum (10%v/v, FBS, R&D systems, cat# S11150) and
penicillin/streptomycin (1%v/v, P/S, Thermofisher, cat# 15070-063) was
used for expansion of HepG2 cells. For HSC cells, High glucose DMEM
containing L-glutamine, sodium pyruvate, sodium bicarbonate (Sigma
Aldrich, cat#D6029), 10% v/v FBS and 1% v/v P/S was used. The
fabrication of cell spheroids with MPs utilized previous
findings.[16] Briefly, HepG2 and HSC cells were
mixed at a 4:1 ratio, then added to MPs to achieve a desired final cell
to particle ratio (150:1) and transferred to the 96-well cell repellant
plates (CELLSTAR® Plates with Cell-Repellent Surface, cat#: 650970) at
a seeding density of 20,000 cells/well. Cells were overlaid with a
serum-free and defined hepatocyte culture media (Corning®, cat#
355056). To form cell aggregates, the well plate was centrifuged at 300
× g for 5 min.