Long Term Model of Human Gut
The FiberCell hollow fibre system has been used to construct a long-term human gut model capable of generating 1E+7 Cryptosporidium parvum oocytes per week. These are the key features:
- An intestinal epithelial cell layer (HCT-8) cultured on the extra-capillary surface of the fibers with aerobic nutrient supply
- A separate anaerobic nutrient rich medium to simulate the gut lumen.
- Intestinal epithelial cells differentiate into crypt-villus units covered with microvilli
- The fiber surface area (2100 cm2) provides efficient nutrient, metabolite and gas exchange between the cell layer and the re-circulating medium.
SEM images of HCT-8 cells cultured in HFBR
Images reproduced here with kind permission of Professor Nigel Yarlett, Director of Haskins Laboratories, Pace University, New York, USA.
Long-term in vitro Culture of Cryptosporidium parvum and use of model in development of therapeutics
References:
In Vitro Culture of Cryptosporidium parvum Using Hollow Fiber Bioreactor: Applications for Simultaneous Pharmacokinetic and Pharmacodynamic Evaluation of Test Compounds:Yarlett, N. et al, Methods Mol Biol 2020 [Abstract]
Yarlett, N.; BIO-PROTOCOL [Online Protocol]
P-glycoprotein mediated efflux reduces the in vivo efficacy of a therapeutic targeting the gastrointestinal parasite Cryptosporidium: Arnold SLM et al; J Infect Dis. 2019 Jun 8. [open access]
Continuous culture of Cryptosporidium parvum using hollow fiber technology: Morada, M. et al.; Int J Parasitol. 2016 Jan;46(1):21-9 [related presentation PDF]
After 8 weeks of growth, the cartridge was cut open and slices of thin sections of the hollow fibers examined by Scanning Electron Microscopy. The intestinal epithelial cells had differentiated into crypt-villus units with microvilli (MV) covering them.
(A) Proximal end of the cartridge shows many craters (C) in the epithelial cell layer, typical of a C. parvum infection.
(B) Distal end of the cartridge has less epithelial damage and developmental stages are visible as sacs covering the villi (DS).