title: A scaffoldless technique for self-generation of three-dimensional keratinospheroids on liquid crystal surfaces.
abstract: We describe a new scaffold-free three-dimensional (3D) cell culture model using cholesteryl ester based lyotropic liquid crystal (LC) substrates. Keratinocytes were deposited randomly on the LC surface where they self-assembled into 3D microtissues or keratinospheroids. The cell density required to form spheroids was optimized. We investigated cell viability using dead/live cell assays. The adhesion characteristics of cells within the microtissues were determined using histological sectioning and immunofluorescence staining. Fourier transform infrared spectroscopy (FTIR) was used to characterize the biochemistry of the keratinospheroids. We found that both cells and microtissues could migrate on the LC surface. The viability study indicated approximately 80% viability of cells in the microtissues up to 20 days of culture. Strong intercellular adhesion was observed in the stratification of the multi-layered microspheroids using field emission-scanning electron microscopy (FE-SEM) and histochemical staining. The cytoskeleton and vinculins of the cells in the microtissues were expressed diffusely, but the microtissues were enriched with lipids and nucleic acids, which indicates close resemblance to the conditions in vivo. The basic 3D culture model based on LC may be used for cell and microtissue migration studies in response to cytochemical treatment.
authors.affiliation: a Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia , Parit Raja, Batu Pahat, Johor , Malaysia.|b Biosensor and Bioengineering Laboratory, MiNT-SRC, Universiti Tun Hussein Onn Malaysia , Parit Raja, Batu Pahat, Johor , Malaysia.
authors.forename: C F
authors.initials: CF
authors.lastname: Soon
authors.affiliation: a Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia , Parit Raja, Batu Pahat, Johor , Malaysia.
authors.forename: K T
authors.initials: KT
authors.lastname: Thong
authors.affiliation: a Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia , Parit Raja, Batu Pahat, Johor , Malaysia.
authors.forename: K S
authors.initials: KS
authors.lastname: Tee
authors.affiliation: b Biosensor and Bioengineering Laboratory, MiNT-SRC, Universiti Tun Hussein Onn Malaysia , Parit Raja, Batu Pahat, Johor , Malaysia.
authors.forename: A B
authors.initials: AB
authors.lastname: Ismail
authors.affiliation: c School of Medical Sciences, University of Bradford , Bradford , United Kingdom.
authors.forename: M
authors.initials: M
authors.lastname: Denyer
authors.affiliation: a Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia , Parit Raja, Batu Pahat, Johor , Malaysia.
authors.forename: M K
authors.initials: MK
authors.lastname: Ahmad
authors.affiliation: d Cancer Research Malaysia, Subang Jaya , Selangor , Malaysia.
authors.forename: Y H
authors.initials: YH
authors.lastname: Kong
authors.affiliation: d Cancer Research Malaysia, Subang Jaya , Selangor , Malaysia.
authors.forename: P
authors.initials: P
authors.lastname: Vyomesh
authors.affiliation: d Cancer Research Malaysia, Subang Jaya , Selangor , Malaysia.
authors.forename: S C
authors.initials: SC
authors.lastname: Cheong
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002470:Cell Survival; D005455:Fluorescent Antibody Technique; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D050866:Liquid Crystals; D017550:Spectroscopy, Fourier Transform Infrared; D023822:Tissue Engineering
keywords: Fourier transform infrared spectrometry; aggregates; cell culture; cell viability; cholesteric liquid crystal; cytoskeleton; keratinocytes; keratinospheroids; live/dead cell assay; microtissues; scaffolds; self-assembly; self-generation; spheroids; three-dimensional cell culture