Gradient Perfusion Culture - Simulating a Tissue-Specific Environment for Epithelia in Biomedicine

Abstract

Epithelia act in the organism as biological barriers. All of them are exposed to different environments at the luminal and basal side. To simulate such a tissue-specific situation Minusheet ® gradient perfusion culture was developed. For pharmaceutical research, biomaterial testing and tissue engineering epithelial cells are cultured on individually selected supports (1). Growing epithelia are stabilized within a tissue carrier (2). Long term culture is performed in a gradient perfusion container (3). To expose epithelia to a tissue-specific environment fresh media of different composition are transported parallel to the luminal and basal compartment of the gradient container. During culture leakage, edge damage and pressure differences have to be avoided. Harvest of intact epithelia is promoted by the use of biocompatible supports and innovative equipment such as transport of oxygen-rich and gas bubble-free medium. Actual literature demonstrates that gradient perfusion culture is an effective method to investigate barrier functions under realistic conditions. Examples of application comprise renal epithelia, retina, blood-air barrier, blood-brain barrier including aspects of tissue-specific development and regeneration.

Zidovudine and Ursodeoxycholic Acid Conjugation: Design of a New Prodrug Potentially Able To Bypass the Active Efflux Transport Systems of the Central Nervous System

“…57 To improve the culture environment for the HRPE epithelial cell line, Millicell filter inserts were applied, feeding the HRPE cell monolayer with media of different composition at the apical and basolateral side. 58 This arrangement allowed the cells to acquire directional polarization characterized by epithelial barrier properties, as demonstrated by TEER measurements (about 80 Ω•cm 2 ). After confluence, the HRPE cell layer separated an upper from a lower compartment.…”

Bridging the gap between traditional cell cultures and bioreactors applied in regenerative medicine: practical experiences with the MINUSHEET perfusion culture system

“…Application of a gradient perfusion culture container made it further possible to investigate the influence on differentiation of different fluid composition at the luminal and basal sides of embryonic renal collecting duct (CD) epithelia (Minuth et al 1992b(Minuth et al , 1997a(Minuth et al , b, 1999(Minuth et al , 2001(Minuth et al , 2005aSteiner et al 1997, Schumacher et al 2002aMinuth et al 2009a). In the course of performed experiments it was detected that development of a CD epithelium starts with an unexpected long latent period of three days and needs at least 10 days for up-regulation of typical signs of differentiation.…”

The Use of Molecularly Imprinted Polymers for Dermal Drug Delivery

“…The alternative advantage of these biological mimicking polymers provides insights into understanding the biofunctional and self-organization within living tissues that will be valuable for clinical implications. There has been a plethora of experiments to determine percutaneous absorption in vitro and it is important to select a design where the transport is limited by the skin and not in any stagnant diffusion layer placed to the underlying skin surface, and which this is related to bio membrane research [71,72]. The applicability of composite, functionalized MIP has shown to be advantageous to the elucidation of the normal functioning of the skin when the template enantiomer molecule binds to the respective MIP, the active triggers the scintillator of living cells to generate a response signal [7,73].…”