Silica bioreplication preserves three-dimensional spheroid structures of human pluripotent stem cells and HepG2 cells.
Three-dimensional (3D) cell cultures produce more in vivo-like multicellular structures such as spheroids that cannot be obtained in two-dimensional (2D) cell cultures. Thus, they are increasingly employed as models for cancer and drug research, as well as tissue engineering. It has proven challenging to stabilize spheroid architectures for detailed morphological examination. Here we overcome this issue using a silica bioreplication (SBR) process employed on spheroids formed from human pluripotent stem cells (hPSCs) and hepatocellular carcinoma HepG2 cells cultured in the nanofibrillar cellulose (NFC) hydrogel. The cells in the spheroids are more round and tightly interacting with each other than those in 2D cultures, and they develop microvilli-like structures on the cell membranes as seen in 2D cultures. Furthermore, SBR preserves extracellular matrix-like materials and cellular proteins. These findings provide the first evidence of intact hPSC spheroid architectures and similar fine structures to 2D-cultured cells, providing a pathway to enable our understanding of morphogenesis in 3D cultures.
Centre for Drug Research, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, the University of Helsinki, Helsinki 00014, Finland.
Yan-Ru
YR
Lou
Centre for Drug Research, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, the University of Helsinki, Helsinki 00014, Finland.
Liisa
L
Kanninen
Advanced Materials Laboratory, Sandia National Laboratories, Albuquerque, New Mexico 87185, USA.|Department of Chemical and Biomolecular Engineering, the University of New Mexico, Albuquerque, New Mexico 87131, USA.
Bryan
B
Kaehr
Division of Molecular Medicine, Department of Internal Medicine, the University of New Mexico, Albuquerque, New Mexico 87131, USA.|Center for Micro-Engineered Materials, the University of New Mexico, Albuquerque, New Mexico 87131, USA.
Jason L
JL
Townson
Centre for Drug Research, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, the University of Helsinki, Helsinki 00014, Finland.
Johanna
J
Niklander
Centre for Drug Research, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, the University of Helsinki, Helsinki 00014, Finland.
Riina
R
Harjumäki
Advanced Materials Laboratory, Sandia National Laboratories, Albuquerque, New Mexico 87185, USA.|Department of Chemical and Biomolecular Engineering, the University of New Mexico, Albuquerque, New Mexico 87131, USA.
C
C
Jeffrey Brinker
Centre for Drug Research, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, the University of Helsinki, Helsinki 00014, Finland.
Marjo
M
Yliperttula
D018929:Cell Culture Techniques; D056945:Hep G2 Cells / Q000166:cytology; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D007150:Immunohistochemistry; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology; D012822:Silicon Dioxide / Q000737:chemistry*; D018874:Spheroids, Cellular / Q000166:cytology*
D020100:Hydrogels; D012822:Silicon Dioxide