Rapid cell sheet detachment from Poly(N-isopropylacrylamide)-grafted porous cell culture membranes

Abstract

Fabrication of functional tissue constructs using sandwiched layers of cultured cells could prove to be an attractive approach to tissue engineering. Rapid detachment of cultured cell sheets is a very important recovery method that permits facile manipulation of the sheet and prevents functional damage. To accelerate the required culture substrate hydrophilic and hydrophobic structural changes in response to culture temperature alteration, poly(N-isopropylacrylamide) (PIPAAm) was grafted onto porous culture membranes by electron beam irradiation. Analyses by attenuated total reflection-Fourier transform IR and electron spectroscopy for chemical analysis revealed that PIPAAm was successfully grafted to surfaces of porous membranes. Atomic force microscopy (AFM) results showed that PIPAAm-grafted membranes had smoother surfaces than ungrafted controls while retaining their porous structure. The mean roughness of PIPAAm-grafted and -ungrafted porous membrane surfaces determined by digital AFM autocalculation was 4.40 +/- 0.4 and 5.9 +/- 0.4 nm, respectively. Tissue culture polystyrene (TCPS) dishes grafted with PIPAAm were compared with PIPAAm-grafted porous membranes in cell sheet detachment experiments. Approximately 75 min was required to completely detach cell sheets from PIPAAm-grafted TCPS surfaces compared to only 30 min to detach cell sheets from PIPAAm-grafted porous membranes. With porous membranes, the water accesses the PIPAAm-grafted surface from underneath and peripheral to the attached cell sheet, resulting in rapid hydration of grafted PIPAAm molecules and detachment of the cell sheet. With TCPS PIPAAm-grafted surfaces the water is supplied from only the periphery of a cell sheet, slowing detachment.

Predicting biomaterial property-dendritic cell phenotype relationships from the multivariate analysis of responses to polymethacrylates

“…Therefore, the conclusion drawn in the PCA model is only valid for this range of contact angles. The higher contact angles (close to 71.2°) are similar to that of TCPS [39], which is known to be suitable for most cell cultures and may be approximately the optimal contact angles for maintaining DC immature state. However, close inspection of Table 3 indicated that no strong correlation exists between DC maturation and contact angle, but pMAs with higher contact angles do group towards the left of Table 3 with pMAs such as pHEMA and pHPMA that caused low DC maturation.…”

Cell sheet tissue engineering: Cell sheet preparation, harvesting/manipulation, and transplantation

“…6(B)]. 64 To this end, we grafted PIPAAm on a porous PET membrane by electron beam-irradiated polymerization of IPAAm. The graft density of PIPAAm was 1.4 μg/cm 2 on porous membrane while that on the control TCPS surface was 1.6 μg/cm 2 , demonstrating that almost identical graft density could be obtained.…”

Thin Hydrogel Films for Optical Biosensor Applications

“…Their ability to contain water, stability in aqueous media and softness, makes hydrogels compatible with biological systems and thus they were proposed for different biomedical applications such as tissue engineering, cell adhesion, wound healing, controlled drug release, contact lenses, to name a few [3,4,5,6,7,8,9,10,11]. Hydrophilic polymer layers were also employed as support for lipid bilayer membranes, in particular for electric measurements of transport phenomena involving membrane proteins [12,13,14,15,16].…”