Download Del “Hígado en un Chip” al hígado bioartificial.

CURSO DE HEPATOLOGÍA BÁSICA Y TRANSLACIONAL:
“Bioingeniería aplicada a la hepatología”
Del “Hígado en un Chip” al
hígado bioartificial.
Rosa Villa
Grupo Aplicaciones Biomédicas (IMB-­CNM, CSIC)
Del hígado en un chip al hígado bioartificial
Multiscale tissue engineering for liver reconstruction
Organogenesis.2014
R Sudo. Keio University;; Yokohama, Japan
Del hígado en un chip al hígado bioartificial
MICROELECTRONIC TECHNOLOGIES
Del hígado en un chip al hígado bioartificial
LAB ON A CHIP
MICROFLUIDICS DEVICES, MICRO TOTAL ANALYSIS SYSTEMS (µ-TAS)
Microfluidic diagnostic technologies for global public health.
Nature. 2006 Jul
Yager et als. University of Washington
Del hígado en un chip al hígado bioartificial
ORGAN ON A CHIP
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q The convergence of Lab-on-Chips and cell biology has
permitted the study of human physiology in an organ-specific
context, introducing a novel model of in vitro multicellular
human organisms.
q An Organ-on-a-Chip is a multi-channel 3-D microfluidic cell
culture chip that simulates the activities, mechanics and
physiological response of entire organs and organ systems.
Del hígado en un chip al hígado bioartificial
ORGAN ON A CHIP
Lung on chip. Huh et al., Nature Protocols 2012
Del hígado en un chip al hígado bioartificial
ORGAN ON A CHIP
Microfluidic devices have been developed to investigate the biological responses of cells and tissues to various mechanical stimuli. Microfluidic platforms for mechanobiology. Lab on a chip. 2013
Polacheck WJ. Massachusetts Institute of Technology
Del hígado en un chip al hígado bioartificial
LIVER ON A CHIP
Adams et al. Nature Reviews Immunology 6, 244–251 (March 2006) |
0.1-­‐ 0.5 dyne/cm2 A biologically inspired liver sinusoid
with a microfluidic endothelial-­‐like
barrier (a) A SEM micrograph
depicts the microflui dic sinus oid
unit. (b) A cord of hepatocytes in
the cell area region of the device is
fed
nutrients
across
the
endothelial-­‐like
barrier
via
convective flow as media is fed
through the outer flow channels.
This device sustained rat and
human hepatocytes for 7 days
without
the introduction of
exogenous matrix
Architecture of Liver Sinusoid.
The 3-D BIOMIMETIC LIVER Device
Physiologies to Captute
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•
•
•
•
Cellular Mechanisms of Action
Albumin, Urea, LDH leakage, Glucosa
Drug metabolism
Zonation (O2, chemical)
Bile Production
(a) Results of cell alignment and culture. a Aligned cells in two lines like a hepatic cord.
(b) Bile canaliculi formation in the cell culture area. Hepatocytes formed bile canaliculi
along the hepatic cord-­‐like structure.
(c) Control experiment in a 24 well plate. Bile canaliculi were formed randomly in the well plate.
LP Lee
Bile canaliculi formation by aligning rat primary hepatocytes in a microfluidic device
Biomicrofluidics 5, 022212 (2011)
Del hígado en un chip al hígado bioartificial
LIVER ON A CHIP
Identification of the four types of mouse hepatic
cells in an in vitro 3D liver sinusoid liver chip
cultured for 1 day. A–D. Immunostaining of
respective biomarkers for the distinct types of
cells. Hepatocytes stained with E-­‐cadherin (red),
CK pan (green) and nucleus (blue ) are closely
seeded in the lower channel of the assemble d
liver chip.
Intriguingly, co-­‐culture and shear flow enhance albumin secretion independently or
cooperatively, while shear flow alone enhances HGF production and CYP450 metabolism.
Mimicking liver sinusoidal structures and functions using a 3D-­configured microfluidic chip. Lab Chip 2017
M. Long et als. University of Chinese
Del hígado en un chip al hígado bioartificial
X
Retina on a chip Biomedical Applications Group
Del hígado en un chip al hígado bioartificial
2015
2016-­‐2017
Patent: EP14157145.5
lla X, Vila S, Yeste J, Peralta C, Gracia-­Sancho J, Villa R
A Novel Modular Bioreactor to In Vitro Study the Hepatic Sinusoid
Plos One 9 (2014) e111864
Del hígado en un chip al hígado bioartificial
LIVER ON A CHIP
(a)
(b)
(c)
Excretory function
(d)
(e)
CYP3A4 Activity
MAINTENANCE OF HEPATOCYTE P HENOTYPE IN VITRO
HUMAN 3 -­‐7 DAYS
Gene e xpression (lost prevention) Dr. J. Gracia-Sancho
Liver Vascular Biology Research Group
Gene e xpression (gain prevention) Del hígado en un chip al hígado bioartificial
LIVER ON A CHIP
1.
2.
3.
4.
Drug development and targeting
Human disease modeling
Personalized medicine
Artificial Liver or support
Del hígado en un chip al hígado bioartificial
Del hígado en un chip al hígado bioartificial
Microbioreactor systems
3D printing technologies
The LiverChip® is a multiwell liver-­‐on-­‐a-­‐chi p device
that enables extended (>1 month) maintenan ce of
phenotypically intact three-­‐dimensional primary
human liver cells. It consists of fluidically isolated
wells that house scaffolds for cells to adhere to and
culture in perfused conditions.
Primary rat hepatocytes and stabilizing stromal fibroblasts
in agarose gels. Cells survive at t he gel perimeter and near
perfused channels ( 8 days od culture)
L.G. Griffith
Perfused multiwell plate for 3D liver tissue engineering
Lab Chip, 10 (2010)
S. Bathia
Rapid casting of patterned vascular networks for perfusable engineered 3D tissues. Nat Mater. (2012) . MIT. Microfabricated
hydrogel
s. Takeuchi
Metre-­long cell-­laden microfibres exhibit
tissue morphologies and functions
Nature Materials 12 (2013) Del hígado en un chip al hígado bioartificial
GRACIAS!!
rosa.villa@csic.es