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Engineered Tissues for in vitro Research Uses, Arrays Thereof, and Methods of Making the Same

a technology tissue, applied in the field of in vitro research tissue engineering tissues, arrays thereof, and methods of making the same, can solve the problems of low rate of new therapeutic discovery, unsustainable r&d costs, and the cost of drug discovery is approximately $1.8 billion, so as to increase the number and quality of innovative, cost-effective, and the effect of improving the quality of new therapeutic discovery

Inactive Publication Date: 2018-11-01
ORGANOVO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a method for creating living, three-dimensional tissue constructs that can be used for drug testing and other applications. These constructs are made by combining adherent cells in a 3D printing process. The resulting tissue has a multi-layered architecture and can be used for in vitro assays and drug testing. The method can also be used to create vascular wall segments for use in drug screening. The adherent cells can be derived from various tissues in the body. The tissue constructs can be coated with biocompatible materials and placed onto a porous membrane for further use. Overall, the method allows for the creation of larger, more complex tissue structures with improved precision and efficiency.

Problems solved by technology

The research and development cost of a new pharmaceutical compound is approximately $1.8 billion.
Despite advances in technology and understanding of biological systems, drug discovery is still a lengthy, expensive, and inefficient process with low rate of new therapeutic discovery.

Method used

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  • Engineered Tissues for in vitro Research Uses, Arrays Thereof, and Methods of Making the Same
  • Engineered Tissues for in vitro Research Uses, Arrays Thereof, and Methods of Making the Same
  • Engineered Tissues for in vitro Research Uses, Arrays Thereof, and Methods of Making the Same

Examples

Experimental program
Comparison scheme
Effect test

example 1

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Smooth Muscle Cells

[0194]Primary human aortic smooth muscle cells (HASMC; GIBCO / Invitrogen Corp., Carlsbad, Calif.) were maintained and expanded in low glucose dulbecco's modified eagle medium (DMEM; Invitrogen Corp., Carlsbad, Calif.) supplemented with 10% fetal bovine serum (FBS), 100 U / mL Penicillin, 0.1 mg / mL streptomycin, 0.25μg / mL of amphotericin B, 0.01M of HEPES (all from Invitrogen Corp., Carlsbad, Calif.), 50 mg / L of proline, 50 mg / L of glycine, 20 mg / L of alanine, 50 mg / L of ascorbic acid, and 3 μg / L of CuSO4 (all from Sigma, St. Louis, Mo.) at 37° C. and 5% CO2. Confluent cultures of HASMC between passage 4 and 8 were used in all studies.

Endothelial Cells

[0195]Primary human aortic endothelial cells (HAEC; GIBCO / Invitrogen Corp., Carlsbad, Calif.) were maintained and expanded in Medium 199 (Invitrogen Corp., Carlsbad, Calif.) supplemented with 10% FBS, 1 μg / mL of hydrocortisone, 10 ng / mL of human epidermal growth factor, 3 ng / mL of basic fibroblast growth factor, 10 μg...

example 2

Solutions and Mold

[0200]Preparation of 2% and 4% (w / v) NovoGel™ Solution

[0201]1 g or 2 g (for 2% or 4% respectively) of NovoGel™ (Organovo, San Diego, Calif.) was dissolved in 50 mL of Dulbecco's phosphate buffered saline (DPBS; Invitrogen Corp., Carlsbad, Calif.). Briefly, the DPBS and NovoGel™ are heated to 85° C. on a hot plate with constant stirring until the NovoGel™ dissolves completely. NovoGel™ solution is sterilized by steam sterilization at 125° C. for 25 minutes. The NovoGel™ solution remains in liquid phase as long as the temperature is maintained above 65.5° C. Below this temperature a phase transition occurs, the viscosity of the NovoGel™ solution increases and the NovoGel™ forms a solid gel.

Preparation of NovoGel™ Mold

[0202]An NovoGel™ mold was fabricated for the incubation of cylindrical bio-ink using a Teflon® mold that fit a 10 cm Petri dish. Briefly, the Teflon® mold was pre-sterilized using 70% ethanol solution and subjecting the mold to UV light for 45 minutes. ...

example 3

on of HASMC-HAEC Polytypic Cylindrical Bio-Ink

[0203]To prepare polytypic cylindrical bio-ink, HASMC and HAEC were individually collected and then mixed at pre-determined ratios. Briefly, the culture medium was removed from confluent culture flasks and the cells were washed with DPBS (1 ml / 5 cm2 of growth area). Cells were detached from the surface of the culture flasks by incubation in the presence of trypsin (1 ml / 15 cm2 of growth area; Invitrogen Corp., Carlsbad, Calif.) for 10 minutes. HASMC were detached using 0.15% trypsin while HAEC were detached using 0.1% trypsin. Following the incubation appropriate culture medium was added to the flasks (2× volume with respect to trypsin volume). The cell suspension was centrifuged at 200 g for 6 minutes followed by complete removal of supernatant solution. Cell pellets were resuspended in respective culture medium and counted using a hemocytometer. Appropriate volumes of HASMC and HAEC were combined to yield a polytypic cell suspension co...

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Abstract

Disclosed are living, three-dimensional tissue constructs for in vitro scientific and medical research, arrays thereof, and methods of making said tissues and arrays.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of U.S. application Ser. No. 13 / 612,768, filed Sep. 12, 2012, which claims the benefit of U.S. Application Ser. No. 61 / 533,757, filed Sep. 12, 2011, U.S. Application Ser. No. 61 / 533,753, filed Sep. 12, 2011, and U.S. Application Ser. No. 61 / 533,761, filed Sep. 12, 2011, all of which are hereby incorporated by reference in their entirety.BACKGROUND OF THE INVENTION[0002]The research and development cost of a new pharmaceutical compound is approximately $1.8 billion. See Paul, et al. (2010). How to improve R&D productivity: the pharmaceutical industry's grand challenge. Nature Reviews Drug Discovery 9(3):203-214. Drug discovery is the process by which drugs are discovered and / or designed. The process of drug discovery generally involves at least the steps of: identification of candidates, synthesis, characterization, screening, and assays for therapeutic efficacy. Despite advances in technology and underst...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/50A61L27/50C12N5/071A61L27/34A61L27/38
CPCG01N33/5088C12N2502/27A61L27/50C12N5/0697C12N5/0691A61L27/34C12N2513/00A61L27/3891C12N2502/28G01N33/5082A61L27/38
Inventor MURPHY, KEITHKHATIWALA, CHIRAGDORFMAN, SCOTTSHEPHERD, BENJAMINPRESNELL, SHARONROBBINS, JUSTIN
Owner ORGANOVO
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