title: Perfusion culture enhanced human endometrial stromal cell growth in alginate-multivalent integrin α5β1 ligand scaffolds.
abstract: A method to functionalize alginate by introducing monomeric or self-assembling (tetrameric) fibronectin (FN) domains is described, leading to a functional scaffold, which is used for three dimensional (3D) culture of human endometrial stromal cells (EnSCs). EnSCs encapsulated in the functional alginate were cultured under perfusion using the TissueFlex® platform, a multiple parallel microbioreactor system for 3D cell culture. The effect of the novel scaffold and the effect of perfusion were examined. Cell viability, proliferation, and extracellular matrix (ECM) deposition were determined and the results compared with those obtained with cells encapsulated in non-functionalized alginate, and also those without perfusion. Staining for focal adhesions and actin showed maximal cell adhesion only for alginate-tetrameric FN scaffolds under perfusion, associated with a significant increase in cell number over 7 days culture; in contrast to poor cell adhesion and a decrease in cell number for non-functionalized alginate scaffolds (irrespective of perfused/static culture) and 3D static culture (irrespective of the scaffold). Conjugation of alginate to FN was an absolute requirement to attenuate the loss of cell metabolic activity over 7 days culture. ECM deposition for blank alginate and alginate-monomeric FN was similar, but increased around 2-fold and 3-fold for alginate-tetrameric FN under static and perfusion culture, respectively. It is concluded that the requirement for EnSC engagement with multivalent integrin α5β1 ligands and perfused culture are both essential as a first step toward endometrial tissue engineering.
authors: Nuffield Department of Obstetrics and Gynaecology, University of Oxford, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors: Zhaohui
authors: Z
authors: Li
authors: Michaela
authors: M
authors: Kreiner
authors: RuAngelie
authors: R
authors: Edrada-Ebel
authors: Zhanfeng
authors: Z
authors: Cui
authors: Christopher F
authors: CF
authors: van der Walle
authors: Helen J
authors: HJ
authors: Mardon
mesh_terms: D000199:Actins / Q000378:metabolism; D000464:Alginates / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D019149:Bioreactors; D002236:Carbohydrate Conformation; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D004717:Endometrium / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D022001:Focal Adhesions / Q000378:metabolism; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D039081:Integrin alpha5beta1 / Q000378:metabolism*; D008024:Ligands; D008422:Materials Testing; D051419:Paxillin / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000199:Actins; D000464:Alginates; D020100:Hydrogels; D039081:Integrin alpha5beta1; D008024:Ligands; D051419:Paxillin