Use of hydrogel scaffolds to develop an in vitro 3D culture model of human intestinal epithelium

Abstract

Christine (2017). Use of hydrogel scaffolds to develop an in vitro 3D culture model of human intestinal epithelium. Acta biomaterialia. Copyright and re-use policy

Developing models of the small intestine

“…This was seen both in Caco-2 monoculture after 3 weeks and in co-cultures where differentiation had taken place. Furthermore, in monoculture and co-culture models no loss of metabolic activity was observed, which may be due to increased flow of nutrients and oxygen by the use of dynamic culture conditions, this effect has been previously seen in Chapter 3 when cells are grown under dynamic culture conditions (Dosh et al 2017). For long-term co-culture experiments, the optimal seeding percentages were 90% Caco-2 / 10% HT29-MTX cells and 75% Caco-2 / 25% HT29-MTX cells.…”

“…The previous study 139 described in Chapter 3 established 3D in vitro culture models of the intestinal epithelium consisting of monocultures of Caco-2 and HT29-MTX cells layered on L-pNIPAM hydrogel scaffolds under dynamic culture conditions. This model supported the 3D culture of these cells generating villus-like structures and promoted differentiation, mimicking the native intestinal epithelium (Dosh et al 2017). Following gelation 300 µL or 100 µL of 2x10 6 total cells/ml (for each of the 6 cell culture suspensions) in complete media were applied to the surface of hydrogel construct in 48 and 96 well plates respectively, and following a 30 minute cell attachment period a further 200 µL or 150 µL complete media (Caco-2 culture media) was added to each well.…”

“…However, these fabricated scaffolds did not form true-villi, but the scaffolds moulded to form villi like structures for cells to grow on. In contrast co-cultured human colon carcinoma cell lines Caco-2 and HT29-MTX cells grown on L-pNIPAM have been shown to spontaneous give rise to villus-like structures (Dosh et al 2017). In the current study, cultures demonstrated the early stages of organoid formation and cellular differentiation.…”

“…This study demonstrated that a synthetic extracellular matrix could provide a suitable scaffold for maintenance and expansion of crypts and intestinal stem cells. Thus, the current study investigated the potential of the novel synthetic L-pNIPAM hydrogel, (which have previously demonstrated the ability to support villus like structures formation in Caco-2 cells and HT29-MTX cells (Dosh et al 2017) as a 3D scaffold for crypt cells and to support long-term culture of Lgr5 + intestinal stem cells under dynamic culture conditions. In addition, the capacity of these cells to differentiate into intestinal epithelial cells in vitro when suspended within or layered on L-pNIPAM hydrogel were also investigated.…”

Recent Strategies in Fabrication of Gradient Hydrogels for Tissue Engineering Applications

“…For example, Kim et al demonstrated that bone morphogenetic protein‐2 (BMP‐2) incorporated thiolated gelatin and poly(ethylene glycol) diacrylate (PEGDA) composite hydrogel scaffold could exhibit effective regeneration of calvarial bone in a rat model 29. Dosh et al utilized poly( N ‐isopropylacrylamide) (PNIPAM) hydrogel as an in vitro 3D culture model for human intestinal cells and suggested that the hydrogel model could be a potential 3D model for intestinal tissue regeneration 30. However, conventional approaches of homogeneous artificial environments have limitations in completely mimicking the natural cellular environment 31…”