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

Abstract

To meet specific requirements of developing tissues urgently needed in tissue engineering, biomaterial research and drug toxicity testing, a versatile perfusion culture system was developed. First an individual biomaterial is selected and then mounted in a MINUSHEET Ò tissue carrier. After sterilization the assembly is transferred by fine forceps to a 24 well culture plate for seeding cells or mounting tissue on it. To support spatial (3D) development a carrier can be placed in various types of perfusion culture containers. In the basic version a constant flow of culture medium provides contained tissue with always fresh nutrition and respiratory gas. For example, epithelia can be transferred to a gradient container, where they are exposed to different fluids at the luminal and basal side. To observe development of tissue under the microscope, in a different type of container a transparent lid and base are integrated. Finally, stem/progenitor cells are incubated in a container filled by an artificial interstitium to support spatial development. In the past years the described system was applied in numerous own and external investigations. To present an actual overview of resulting experimental data, the present paper was written.

Apical Medium Flow Influences the Morphology and Physiology of Human Proximal Tubular Cells in a Microphysiological System

“…Indeed, the apical epithelial surface in the kidney proximal tubule is exposed to the shear stress generated by the constant flow of the glomerular filtrate in vivo. Systems able to generate perfusion cultures applied to the kidney cell cultures include the work by Minuth [ 11 ], Ferrell [ 12 ], and Brakeman [ 13 ]. Accordingly, Duan et al demonstrated that fluid shear stress (FSS)-induced actin cytoskeletal reorganization and junctional formation in renal epithelial cells and altered expression of apical and basolateral transporters in cultured mouse proximal tubule cells [ 14 , 15 ].…”

In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers

“…To improve the environment in which cells and developing tissues can be maintained under in vitro conditions, the MINUSHEET perfusion culture system was developed. In this perfusion system, a constant flow of culture medium provides seeding cells and tissues with a constant supply of fresh nutrients and respiratory gases, making it possible to produce a variety of specialized tissues of the high cell biological quality that is urgently needed in tissue engineering 18 . The use of a perfusion system benefits cells by providing a continuous supply of nutrients and constant removal of metabolic waste.…”

“…Flow perfusion systems provide a flexible platform for developing a controllable biomimetic environment that can be adapted for use in various investigations of dynamic cell culture under conditions of rheological stress and hydrostatic pressure 17 – 19 . Flow perfusion systems such as bioreactors enable the continuous and constant supply of nutrients and oxygen to cells and can be used to improve the environment for in vitro tissue-engineered conditions 18 , 20 . Therefore, flow perfusion systems may be an efficient method for the construction of CEC layers and the observation of morphological and physiological changes in CECs.…”

“…The development of a gingival epithelium or co-culture of keratinocytes and osteoblast-like cells in a perfusion container yields much better results than those obtained under static culture conditions. The MINUSHEET perfusion culture system thus is advantageous for use in the engineering of connective tissue, the generation of nervous tissue, and the development of muscle tissue 18 .…”

A bioartificial environment for kidney epithelial cells based on a supramolecular polymer basement membrane mimic and an organotypical culture system

“…Only a limited number of bioreactors that provide a culture environment suitable for renal epithelial cells are commercially available. The ones that allow a free choice of support material and application of flow at each membrane side are, to our knowledge, limited to the culture systems of Minucells and Minutissue GmbH (Minuth et al , ; Minuth and Denk, ). However, none of these systems meet the exact requirements that are desired for the cell studies on our UPy‐based BM mimics.…”