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Engineered three-dimensional skin tissues, arrays thereof, and methods of making the same

Inactive Publication Date: 2016-05-05
ORGANOVO +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes how to create a 3D model of human skin that contains all the important layers and cells. These models can be used instead of animal models in pharmaceutical and cosmetic industries, and can also be used for therapeutic purposes. The advantage of using these 3D models is that they are more physiologically relevant than traditional 2D models, as cells in 3D conformation may behave differently than cells in a 2D monolayer. This patent is useful for anyone working in the field of skin research or development.

Problems solved by technology

Further, complexity is added by the addition of a hypodermal compartment comprising endothelial cells.
In certain embodiments, the method comprises a temporal delay in the deposition of the epidermal bio-ink onto the dermal bio-ink.

Method used

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  • Engineered three-dimensional skin tissues, arrays thereof, and methods of making the same
  • Engineered three-dimensional skin tissues, arrays thereof, and methods of making the same
  • Engineered three-dimensional skin tissues, arrays thereof, and methods of making the same

Examples

Experimental program
Comparison scheme
Effect test

example 1

Bioprinting Full Thickness Skin Tissue by Continuous Deposition Using Dermal Bio-Ink Containing Alginate and Epidermal Bio-Ink Deposition by Aerosol Spray Method

Procedures

[0175]Bio-ink was generated by a cellular mixture of 100% primary adult human dermal fibroblasts (HDFa) in 6% gelatin and 1% alginate (Novogel® 3.0) in a concentration of 150 million cells per milliliter. Three-dimensional bio-ink constructs were printed by continuous deposition using the Novogen Bioprinter® platform in a 4 mm×4 mm×0.5 mm base sheet with a 1 mm wall bordering the top to create a dermal structure resembling a cup. One tissue construct was printed per transwell in a 6 well plate. The transwell printing surface contained a polytetrafluoroethylene (PTFE) membrane coated with equimolar mixture of types I and III collagen (bovine) with pores 3 μm in size. Following printing, constructs were immediately cross-linked by submerging in 5 ml of 50 mM calcium chloride for 2-5 minutes. Calcium chloride was then...

example 2

Bioprinting Full Thickness Skin Tissue by Continuous Deposition Using Dermal Bio-Ink Containing Gelatin and Epidermal Bio-Ink Containing Cell Paste

Procedures

[0177]Bio-ink was generated by a cellular mixture of 100% primary adult human dermal fibroblasts (HDFa) in 6% gelatin (Novogel®) in a concentration of 150 million cells per milliliter. Three-dimensional bio-ink constructs were printed by continuous deposition using the Novogen Bioprinter platform in a 4 mm×4 mm×0.5 mm base sheet with a 1 mm wall bordering the top to create a dermal structure resembling a cup. One tissue construct was printed per transwell in a 6 well plate. The transwell printing surface contained a polytetrafluoroethylene (PTFE) membrane coated with equimolar mixture of types I and III collagen (bovine) with pores 3 μm in size. Epidermal cell paste containing a mixture of 95% primary adult human epidermal keratinocytes (HEKa) and 5% primary adult human epidermal melanocytes (HEMa) was then printed on top of the...

example 3

Additional Example of Bioprinting Full Thickness Skin Tissue by Continuous Deposition Using Dermal Bio-Ink Containing Gelatin and Epidermal Bio-Ink Containing Cell Paste

Procedures

[0179]Bio-ink was generated by a cellular mixture of 100% primary adult human dermal fibroblasts (HDFa) in 8% gelatin (Novogel®) in a concentration of 100 million cells per milliliter. The cell: gelatin ratio was altered to reduce the cellular density of the dermal sheet to better mimic dermal tissue in native skin. Three-dimensional bio-ink constructs were printed by continuous deposition using the Novogen Bioprinter® platform in a 4 mm×4 mm×0.5 mm base sheet to create a dermal structure resembling a sheet. One tissue construct was printed per transwell-in a 6 well plate. The transwell printing surface contained a polytetrafluoroethylene (PTFE)-membrane coated with equimolar mixture of types I and III collagen (bovine) with pores 3 μm in-size. Epidermal cell paste containing a mixture of 100% primary neona...

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Abstract

Disclosed are bioprinted, three-dimensional, biological skin tissues comprising: a dermal layer comprising dermal fibroblasts; and an epidermal layer comprising keratinocytes, the epidermal layer in contact with the dermal layer to form the three-dimensional, engineered, biological skin tissue. Also disclosed are arrays of engineered skin tissues and methods of making engineered skin tissues.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Application Ser. No. 62 / 075,703, filed Nov. 5, 2014 and U.S. Application Ser. No. 62 / 140,381, filed Mar. 30, 2015, the entire disclosures of which are hereby incorporated herein by reference.BACKGROUND OF THE INVENTION[0002]The pharmaceutical and cosmetic industries utilize skin models for toxicology screening, barrier function analysis, pigmentation studies, and models for corrosion, irritation, inflammation, and infection.SUMMARY OF THE INVENTION[0003]Three-dimensional (3D) tissue models for human skin are valuable as an alternative to animal models in both the pharmaceutical and cosmetic industries, and clinically as a therapeutic tissue engraftment. The full thickness human skin models described herein contain a dermal compartment including fibroblasts and connective tissue and an epidermal compartment including stratified keratinocytes. Complexity of the model is optionally increased by inc...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0698C12N2502/091C12N2502/094C12N2502/1323C12N2503/06C12N2533/54C12N2533/74C12N2533/90C12N2535/00A61L27/362A61L27/50A61L27/60A61L2300/64G01N33/5044
Inventor RETTING, KELSEY NICOLEO'NEILL, COLIN M.NGUYEN, DEBORAH LYNN GREENEPRESNELL, SHARON C.
Owner ORGANOVO
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