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.

Cryptosporidium life cycle in hollow fiber gut model

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).

Starter kit for producing hollow fibre gut model

Cat.#Description
P3202FiberCell® Systems Duet
C2011High Flux PS Cartridge 20 kd (3000 cm2 – 20 ml)*
A100638mm Reservoir Cap

* Option :
C5011 High Flux cartridge 20 kd (3000 cm2 – 20 ml) is optimized for hybridoma culture, providing higher flow and oxygenation capacity and therefore greater mAb productivity compared to C2011. Important – C5011 consumes more media than C2011 and is the recommended scale-up option for experienced users. Please enquire.

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