Bio-ingénierie hépatique / Bio-Printing du foie : Enjeux et espoirs Jean-Charles Duclos-Vallée Journées du Centre Hépato-Biliaire - JCHB 2019 Hépatologie

1. Bio-printing du foie:
Enjeux et espoirs
Jean-Charles Duclos-Vallée

2. It always seems impossible
until it’s done
Nelson Mandela

3. Du simple au complexe ?

4. Vers la 4ème dimension
2D
3D
4D
2D

5. Drug development is an expensive venture
$3 to $5 billions and 12 to 15 years
90% of withdrawals of drugs are due to toxicity issues
DRUG DISCOVERY
HIGH THROUGHPUT SCREENING
PHYSIOCHEMICAL PROPERTIES
IN VIVO SCREENING
PHASE 1
PHASE 2
IN VITRO SCREENING
PHASE 3
Lead
Identification
Lead
Optimlzation
Human
Clinical Trials
REGISTRATION & MARKET LAUNCH

6. A STRONG PUSH TO MOVE AWAY FROM
ANIMAL MODELS DUE TO ETHICAL
PROBLEMS FOLLOWING THE 3R
APPROACH
= REDUCTION, REFINEMENT AND REPLACEMENT OF ANIMAL STUDIES
ETHICAL ISSUES
AND PRECLINICAL STUDIES
European commission
Bruxells 11 March 2013

7. An history of 40 yrs of culture
La deuxième dimension

8. Fully functional bio-engineered liver graft
External bio-artificial liver
In vitro liver tissue models

10. Taux d’incidence cumulée de greffe et de
décès ou aggravation sur la liste d’attente
de greffe hépatique (1995-2015)
agence-biomedecine.fr

11. Is the Cellular architecture
of the liver complexe ?

12. Transport polarity of human hepatocytes
Godoy et al Arch Toxicol 2013
Is the Cellular architecture
of the liver complexe ?

13. Time line of mouse liver development
Do we need embryologic
development modelisation?

14. Vers la 4ème dimension
3D
4D
2D

15. Vers la 4ème dimension
2D
3D
2D

18. Du simple au complexe ?

19. Use of whole organ decellularization for the
generation of a vascularized liver organoid
Baptista PM et al, Hepatology 2011; 53(2):604-17.

20. Recellularization of the liver
bioscaffold's parenchyma

21. Basic assumptions for
tissue engineering approaches
! Cell need a solid substrate for attachment and proliferation
! Tissue constructs must have an organo-specific shape
! The scaffold serves also as a source of inductive and instructive
signals for cell differentiation, migration, proliferation and
orientation
! The porous structure of a solid scaffold will allow optimal
seeding, tissue construct viability and vascularization
! Properties will be maintained by controlled neomorphogenesis
of parenchymal and stromal tissue synthezised

22. Will it be possible to master
intercellular language ?

23. Liver democraty

24. Effects of nonparenchymal cell types of the liver on parenchymal
hepatocytes through paracrine secretions.
Salman R. Khetani et al. Journal of Laboratory Automation
2015;20:216-250

25. 3D

26. 3D et coculture

27. Advantages and Pitfalls of spheroids
! Hepatocytes in the
spheroids exhibit
prolonged viability
! Express higher levels of
liver-specific functions as
compared to hepatocyte
monalyer culture
! A more cuboidal
morphology and form
extensive cell-cell contacts
Lu et al. Biomaterials 2003
Yin et al. J Biomed Mater Res 2003
Peshwa et al. In vitro Cell Dev Biol Anim 1996
Antoni D Int J Mol Sci 2014
Usta OB et al. Technology 2015
! Difficulty in imgagin
! A distribution of nutrients,
wastes and test
compounds across the
aggregates
! Difficulty of scaling the
system down for
microfluidic applications

28. hepatocyte
Cholangiocyte

29. Functional properties of hepatic cells generated
From iPSC
Dianat et al Hepatology 2014

31. Self-formation of three-dimensional liver bud
from human iPSC in vitro

32. Generation of Liver Bud from pluripotent
stem cells by mimicking early
organogenesis

33. Therapeutic potential of in vitro grown hiPSC-LB
by mesenteric transplantation model.

34. One-step fabrication of an organ-on-a-
chip with spatial heterogeneity using a 3D
bioprinting
Lee H and Cho DW lab on a Chip 2016

39. Vers la 4ème dimension
2D
3D
4D
2D

40. Solid freeform
fabrication for
biomedical applications
Stereolithography
Selective laser
Sintering (SLS) Plotting
(FDM/3DF)
3D printing
Principle: photopolymerisation
Polymers, ceramics and composites
Hydrogels
cells

41. Determinically patterned biomimetic human iPSC-
derived hepatic model via rapid 3D bioprinting
Ma et al; PNAS 2016
Michels et al Biomaterials 2013

42. Determinically patterned biomimetic human iPSC-
derived hepatic model via rapid 3D bioprinting
Ma et al; PNAS 2016
Michels et al Biomaterials 2013

43. Gammes répétées
Does a simple model exist ?
A repetitive zonation

44. Imaging, image analysis, in silico
models, and software generation
Workflow
Biological question
Data acquisition
(Micrographs, Measurements)
Quantification of tissue
Architecture {P1} & Dynamics {P2}
Validation experiment
Model simulations
(Quantitative) mathematical Model
(implements hypotheses) {P3}
Comparison to experiments
Biological model
(List of hypotheses on mechanisms)
Model adjustments {P3}
Missing agreement
Information missing,
not unique
Published
knowledge
Agreement (Validation)

45. Vers la 4ème dimension
2D
3D
4D
2D

47. un grand principe la bioimpression: la
construction de blocs

48. Vacular block construction

49. Biliary block construction

50. Liver tissue block construction

51. Default of the preexisting models

52. Most commonly used printer types
inkjet Laser-assisted Extrusion
Drop on
demand
Continuous Cell printing
Cell lade in
hydrogels
Cell
patterning
Precision
Low ink waste
Faste
Large scale
Industrial
usage
Low ink waste
No clogging
High resoluion
Expensive
3D constructs
Crucial post-
bioprinting
process

53. Most commonly used printer types
inkjet Laser-assisted Extrusion
Drop on
demand
Continuous Cell printing
Cell lade in
hydrogels
Cell
patterning
Precision
Low ink waste
Faste
Large scale
Industrial
usage
Low ink waste
No clogging
High resoluion
Expensive
3D constructs
Crucial post-
bioprinting
process

55. Un plaidoyer pour la prudence, la démocratie
et l ‘éthique de la discussion
Qu’est ce que le transhumanisme ? Luc Ferry
Association Française Transhumaniste

56. FINANCIAL HIGHLIGHTS
Source: Reportlinker Nov 14, 2016,

57. iLite Project
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