Development of an Evaluation System Using Intestinal Organoids for Drug Efflux Transport Analysis by an Imaging...

Area of Science:

• Pharmacokinetics
• Drug transport
• Intestinal organoids

Background:

• Evaluating drug excretion direction in vitro is challenging.
• Enteroids are intestinal organoids used in research, but the impact of cryopreservation and passaging on transporter function is unclear.

Purpose of the Study:

• To investigate the effects of passage and cryopreservation on enteroid function, specifically P-glycoprotein (P-gp) mediated drug transport.
• To establish an easy and quantitative method for evaluating P-gp substrates using enteroids.

Main Methods:

• Enteroids were subjected to passage and cryopreservation.
• Rhodamine 123 (Rh123) transfer rates into the enteroid lumen were measured with and without P-gp inhibitors.
• mRNA expression levels of transporters were analyzed.

Main Results:

• Passage and cryopreservation did not significantly alter mRNA expression levels of transporters.
• Enteroids demonstrated P-gp-mediated excretion of Rh123 into the lumen.
• P-gp inhibitors significantly reduced Rh123 excretion, leading to intracellular accumulation.
• Rh123 transfer rates remained consistent before and after enteroid manipulation.

Conclusions:

• Enteroids are a reliable model for evaluating P-gp substrates.
• Cryopreservation and passaging do not impair enteroid function for P-gp transport studies.
• This method provides an easy and quantitative way to assess P-gp substrate potential of compounds.