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Copyright 2013, Organovo Holdings, Inc. This report is solely for the use of intended audience.

No part of it may be circulated, quoted, or reproduced for distribution outside the organization without prior written approval from Organovo Holdings, Inc.


This presentation may contain forward-looking statements that involve significant risks and uncertainties. All statements other than statements of historical fact are forwardlooking statements, including statements regarding the Companys prospective performance, opportunities and business outlook. Any forward-looking statements contained herein are based on current expectations, but are subject to a number of risks and uncertainties. The factors that could cause the Companys actual future results to differ materially from current expectations include, but are not limited to, risks and uncertainties relating to the Company's ability to develop, market and sell products based on its technology; the expected benefits and efficacy of the Companys products and technology; the availability of substantial additional funding for the Company to continue its operations and to conduct research and development, clinical studies and future product commercialization; and the Company's business, research, product development, regulatory approval, marketing and distribution plans and strategies. These and other factors are identified and described in more detail in our filings with the SEC, including our annual report for the period ended December 31, 2012 on Form 10K and our current reports filed on Form 8-K. These forward-looking statements are made as of the date of this presentation, and we do not undertake an obligation to update these forward-looking statements after such date. OTC QX: ONVO

Copyright 2013 Organovo, Inc.

Current in vitro cell-based models

are often poor predictors of in vivo outcomes
Attrition rates for experimental and approved drugs remains high
Other; 41% Other 33% Hepatoxicity; Hepatotoxicity 26% 27%

Cardio- and hepatotoxicity are the primary causes for late-stage failures and post-market withdrawals

CV Toxicity; 33% Cardiovascular Toxicity 40%

More robust human models of these organ systems are needed

Drugs withdrawn from global market due to toxicity 1990-2010 (n=39)

From: EvaluatePharma; CDER; Tufts Center for Drug Discovery

Copyright 2013 Organovo Holdings, Inc.

Organovos NovoGen BioprinterTM platform

was utilized to generate a 3D human liver system
Design concept for 3D human liver focused on the following key features: True three-dimensionality, reaching at least 250 microns in the smallest dimension Incorporation of multiple cell types with spatially-controlled placement in x, y, and z axes Demonstration of both histologic and functional features of liver

Copyright 2013 Organovo Holdings, Inc.

Materials and Methods

utilized in these studies
Cell Culture. All cells utilized in these experiments were sourced from commercial vendors and cultured according to manufacturers recommended protocols. Cryo-preserved primary human hepatocytes were purified by Percoll gradient centrifugation prior to use in bioprinted constructs. iPSC-derived hepatocyte-like cells (iCells) were generously provided by Cellular Dynamics, Inc. Bio-ink Preparation. Bio-ink was prepared based on protocols and techniques described in Norotte et al., (Biomaterials 30:5910-5917). For preparation of hydrogels containing cells, NovoGel 2.0 (Organovo, Inc.) was prepared according to manufacturers instructions and non-parenchymal cell populations were incorporated prior to bioprinting. Bioprinting. All tissues were fabricated directly into standard tissue culture plates (Corning Transwell) using standard Organovo bioprinting protocols and the NovoGen MMX BioprinterTM or modifications thereof. Secreted protein detection. Spent media was analyzed by commercially available ELISA kits for the following liver proteins: albumin, fibrinogen, and transferrin. Cholesterol biosynthesis was quantified fluorimetrically (Cayman Biochemical). CYP 450 analysis. CYP1A2 and CYP3A4 activities were assessed with the Pro-GloTM CYP450 Assay system (Promega). Bioprinted liver tissues were challenged with either verapamil (10M) or dexamethasone (10M) to stimulate CYP1A2 or CYP3A4 activity. Fold induction was calculated as the increase in activity of the treated samples relative to the non-treated control samples. Histologic analysis. Tissues were fixed in 10% buffered formalin, paraffin-embedded, and subjected to standard histochemical analyses. In some experiments, tissues were snap frozen upon harvest and cryosectioned prior to histologic or immunohistologic analysis.

Copyright 2013 Organovo Holdings, Inc.

3D liver tissues with relevant architectural features

were fabricated successfully with the NovoGen Bioprinter
Single-Unit Geometry Multi-Unit Geometry Multi-Unit Geometry

100 m

100 m

H&E (20x)

E-Cadherin (20x)

Multi-Unit Tissue (post-fusion)

Single- or Multi-unit bioprinted liver tissues were constructed from hepatocytes or hepatocyte-like cells and non-parenchymal cells, and were characterized by a tissuelike cellular density and tight intercellular junctions.

Copyright 2013 Organovo Holdings, Inc.

Bioprinted 3D human liver tissues

are metabolically active with CYP450 induction
Single Unit / 39 hrs Single Unit / 135 hrs

These liver tissues remained stable for 135 hours and retained key liver functions, including inducible CYP1A2 and CYP3A4 activity.
Constructed w/ 1 hepatocytes, hepatic stellate cells, and endothelial cells


Copyright 2013 Organovo Holdings, Inc.

Bioprinted 3D human liver tissues

synthesize cholesterol
Single-Unit Geometry / 48 hrs

Constructed with HepaRG cells, hepatic stellate cells, and endothelial cells

Bioprinted liver tissues possess the ability to synthesize cholesterol, a hallmark liver-associated function. Cholesterol biosynthesis is retained and even increases over time in culture.

Copyright 2013 Organovo Holdings, Inc.

The NovoGen Bioprinter builds complex 3D tissue

with precise deposition of distinct cell populations

2.5x (light microscope image)

100 m

Hepatic stellate cells (CFSE) Hepatocytes (DAPI) Endothelial cells (CTMB)

Bioprinted human liver tissues are characterized by cell-specific compartments that enable reproduction of native tissue-like architectural patterns and the establishment of controlled cell-cell interfaces.
Copyright 2013 Organovo Holdings, Inc.

3D human liver tissues outperform 2D cultures

with respect to liver-specific protein production

100 m

H&E / 20x

100 m

CD31 / 20x

Bioprinted liver constructs, generated from iPSC-derived hepatocyte-like cells (iCells; Cellular Dynamics, Inc.), are well organized, develop microvascular networks over time, and produce significantly more albumin per cell than matched 2D controls.

Copyright 2013 Organovo Holdings, Inc.

Summary of Experimental Observations

3D human liver tissues were fabricated using an automated bioprinting platform to yield distinct geometries which include both simple and repeating functional units Constructs contained heterogenous cellular inputs including hepatocytes, hepatic stellate cells and endothelial cells Bioprinted constructs were organized, dense and viable, exhibiting microarchitecture in a manner approximating in vivo tissues Sustained production of a number of critical functional endpoints has been demonstrated Albumin, fibrinogen, transferrin, cholesterol, CYP activity The platform enabled generation of functional constructs from a variety of cell sources, including primary hepatocytes and stem/progenitor cell-derived hepatocyte-like cells (HepaRG, iCells) Per-cell production of a liver-specific protein Albumin was significantly enhanced in 3D liver constructs vs. matched 2D cultures

Copyright 2013 Organovo Holdings, Inc.


Bioprinting enabled highly reproducible fabrication of architecturally and compositionally defined 3D tissues into standard tissue culture formats Bioprinted 3D liver tissues exhibited several key unique features that remained stable over time
Tissue-like cellular density with high viability and well-organized microarchitecture (microvasculature, tight junctions) H&E / 20x Cell type-specific compartmentalization , with establishment and retention of userdefined spatial localization of parenchymal and non-parenchymal cells Multi-layered architecture ranging from 250-500 microns in thickness

3D liver tissues possessed critical liver functions, including albumin production, cholesterol biosynthesis, fibrinogen and transferrin production, and inducible CYP1A2 and CYP3A4 activities Per-cell production of Albumin by 3D bioprinted tissues was 5-9x greater than matched 2D controls, suggesting superior functional performance in 3D

Copyright 2013 Organovo Holdings, Inc.