GUT FUNCTION ON-A-CHIP

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Horace Simmons
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1 GUT FUNCTION ON-A-CHIP Towards a more physiological human intestinal barrier model MicroNanoConference &14 December 2016 Evita van de Steeg, PhD

ORGAN FUNCTION-ON-A-CHIP AT TNO APPROACH Development of in vitro models with clinically relevant complexity Focus on areas with best match between TNO expertise and market

2 ORGAN FUNCTION-ON-A-CHIP AT TNO APPROACH Development of in vitro models with clinically relevant complexity Focus on areas with best match between TNO expertise and market need: Make use of existing new technologies within AND outside TNO Collaborations with companies, knowledge institutes and authorities AIMS: The 3 Dimensions of Organ-on-a-Chip

3 GUT FUNCTION ON-A-CHIP WHAT ARE WE AIMING FOR? A personalized in vitro human intestinal model that can be used to study (drug) absorption and impact of drugs, nutrition and environment on gut health Important aspects include: Gut permeability (drugs, antigens, toxins, bacteria) Effect of microbiome on gut health/absorption Detection of early immune response

4 GUT FUNCTION ON-A-CHIP APPROACH Gut function on-a-chip ex vivo intestinal tissue model (intestine TM ) In vitro stemcell derived intestinal organoids

InTESTine TM Integrity markers are used in every study [ 3 H]-mannitol/atenolol (paracellular transport route) [ 14 C]-caffeine /antipyrine

5 INTESTINE TM A MEDIUM THROUGHPUT SYSTEM USING EX VIVO INTESTINAL TISSUE Up to 96 incubations/study Feasible with bio-relevant matrices (e.g. FaSSIF/FeSSIF) Tissue from human, pigs, minipigs, rat (colon) can be used Pig: whole GI tract available, study regional differences InTESTine TM Integrity markers are used in every study [ 3 H]-mannitol/atenolol (paracellular transport route) [ 14 C]-caffeine /antipyrine (transcellular transport route) FD4, MW 4000 (tissue integrity marker) Acceptance criteria: P app C/M or A/A > 3 (jejunum) Leakage of FD4 < 1% / h

6 APPLICATION OF HUMAN INTESTINAL TISSUE Human intestinal tissue is made available from local hospital Patients do sign informed consent ex vivo intestinal tissue model (intestine TM ) ERP OoC: gut function on-a-chip

7 EXTEND VIABILITY OF THE TISSUE towards gut on-a-chip ex vivo intestinal tissue model (intestine TM ) ERP OoC: gut function on-a-chip

8 GUT FUNCTION ON-A-CHIP APPROACH Gut function on-a-chip ex vivo intestinal tissue model (intestine TM ) In vitro stem cell derived intestinal organoids

9 In vitro stemcell derived intestinal organoids CULTURING INTESTINAL ORGANOIDS FROM STEM CELLS

10 STEM CELL DERIVED INTESTINAL ORGANOIDS

11 INTESTINAL ORGANOIDS CULTURED AS MONOLAYER Permeable membrane coated with collagen In vitro stemcell derived intestinal organoids lysozyme Chromogranin A Hoechst F-actin

12 INTESTINAL ORGANOIDS CULTURED AS MONOLAYER Permeable membrane coated with collagen In vitro stemcell derived intestinal organoids T E E R m e a s u r e m e n t 2 D c u ltu re d o rg a n o id s w ell 1 T E E R (O h m *c m 2 ) % F D 4 le a k a g e 2.5 F D 4 le a k a g e d a y 3 2 d a y w ell In collaboration with: Dr. JP ten Klooster Dr. S.F.C. Vaessen T im e (d a y s ) W e ll 1 W e ll 2 W e ll 1 W e ll 2

GUT FUNCTION ON-A-CHIP DEVELOPMENTAL PLANS Validation (reproducibility, robustness) of gut-on-chip platforms for drug absorption, intestinal metabolism, hormone release Effect of fluidics on

13 GUT FUNCTION ON-A-CHIP DEVELOPMENTAL PLANS Validation (reproducibility, robustness) of gut-on-chip platforms for drug absorption, intestinal metabolism, hormone release Effect of fluidics on monolayer culturing of intestinal stem cells Culture cells on nanofabricated permeable membrane scaffolds with villi structure Further implement available state-off-the-art read-out technology (AFM, ring resonators) Focus on co-culture with (anaerobic) microbiota and immune cells (PBMC/DC) Ring resonators AFM my microbiome on a chip

resonators Immune cells Towards a personalized and translational in

14 GUT FUNCTION ON-A-CHIP 3 DIMENSIONS Ex vivo tissue Stem cell organoids Chip platform (3D printed) villi scaffolds AFM microbiota Ring resonators Immune cells Towards a personalized and translational in vitro intestinal model enabling good prediction of GI processes and novel therapeutics!

ACKNOWLEDGEMENTS TNO Organ-on-chip team: Ivana Bobeldijk - Pastorava Mariska Gröllers-Mulderij Gwen Grootaers Roeland Hanemaaijer Mehdi Hassan Zade Nadjari Harm Jansen Alie de Kat Angelino -Bart

15 ACKNOWLEDGEMENTS TNO Organ-on-chip team: Ivana Bobeldijk - Pastorava Mariska Gröllers-Mulderij Gwen Grootaers Roeland Hanemaaijer Mehdi Hassan Zade Nadjari Harm Jansen Alie de Kat Angelino -Bart Ingeborg Kooter Robert Ostendorf Evita van de Steeg Lianne Stevens Karin Toet Application of advanced in vitro models for human prediction GUT FUNCTION ON-A-CHIP COLLABORATORS 2-REAL-GUTS - innovation in intestinal models for food research

16 THANK YOU FOR YOUR ATTENTION! Voor meer inspiratie: TIME.TNO.NL

Effect of sow antibiotic treatment and offspring diet on microbiota and gut barrier throughout life

EAAP Meeting, Nantes, August 29, 2013 Interplay EU Project Workshop 2 Effect of sow antibiotic treatment and offspring diet on microbiota and gut barrier throughout life J.P. Lallès 1, M.E. Arnal 1, G.