title: Integration of hollow fiber membranes improves nutrient supply in three-dimensional tissue constructs.
abstract: Sufficient nutrient and oxygen transport is a potent modulator of cell proliferation in in vitro tissue-engineered constructs. The lack of oxygen and culture medium can create a potentially lethal environment and limit cellular metabolic activity and growth. Diffusion through scaffold and multi-cellular tissue typically limits transport in vitro, leading to potential hypoxic regions and reduction in the viable tissue thickness. For the in vitro generation of clinically relevant tissue-engineered grafts, current nutrient diffusion limitations should be addressed. Major approaches to overcoming these include culture with bioreactors, scaffolds with artificial microvasculature, oxygen carriers and pre-vascularization of the engineered tissues. This study focuses on the development and utilization of a new perfusion culture system to provide adequate nutrient delivery to cells within large three-dimensional (3D) scaffolds. Perfusion of oxygenated culture medium through porous hollow fiber (HF) integrated within 3D free form fabricated (FFF) scaffolds is proposed. Mouse pre-myoblast (C2C12) cells cultured on scaffolds of poly(ethylene-oxide-terephthalate)-poly(butylene-terephthalate) block copolymer (300PEOT55PBT45) integrated with porous HF membranes of modified poly(ether-sulfone) (mPES, Gambro GmbH) is used as a model system. Various parameters such as fiber transport properties, fiber spacing within a scaffold and medium flow conditions are optimized. The results show that four HF membranes integrated with the scaffold significantly improve the cell density and cell distribution. This study provides a basis for the development of a new HF perfusion culture methodology to overcome the limitations of nutrient diffusion in the culture of large 3D tissue constructs.
authors.affiliation: MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, The Netherlands.
authors.forename: N M S
authors.initials: NM
authors.lastname: Bettahalli
authors.forename: J
authors.initials: J
authors.lastname: Vicente
authors.forename: L
authors.initials: L
authors.lastname: Moroni
authors.forename: G A
authors.initials: GA
authors.lastname: Higuera
authors.forename: C A
authors.initials: CA
authors.lastname: van Blitterswijk
authors.forename: M
authors.initials: M
authors.lastname: Wessling
authors.forename: D F
authors.initials: DF
authors.lastname: Stamatialis
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D051379:Mice; D032446:Myoblasts / Q000166:cytology; D010100:Oxygen / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D010100:Oxygen