Integrated 3d cell culture matrix and epithelial support layer

Patent metadata

Field	Value
Publication number	US-2023313104-A1
Application number	US-202318129344-A
Country	US
Kind code	A1
Filing date	Mar 31, 2023
Priority date	Apr 1, 2022
Publication date	Oct 5, 2023
Grant date	—

Abstract

A method for generating a cell support interface for use in a three dimensional cell culture environment, may include electrospinning a mat having an epithelial support layer configured to create an intimate coupling between the epithelial cell and a porous matrix, including a first layer and a second layer, wherein the first layer is formed using a first solution at a first viscosity level and the second layer is formed using a second solution at a second viscosity level different from the first viscosity level.

First claim

What is claimed is:

1 . A method for generating a cell support interface for use in a three dimensional cell culture environment, comprising:
electrospinning a mat having an epithelial support layer configured to create an intimate coupling between the epithelial cell and a porous matrix, including a first layer and a second layer, wherein the first layer is formed using a first solution at a first viscosity level and the second layer is formed using a second solution at a second viscosity level different from the first viscosity level.

2 . The method of claim 1 , wherein the first viscosity level is higher than the second viscosity level to form the first layer at a porosity lower than that of the second layer.

3 . The method of claim 2 , wherein a water-soluble polymer is formed with the second layer, and the method further comprises dissolving the water-soluble polymer in water.

4 . The method of claim 3 , further comprising drying the support layer.

5 . The method of claim 1 , wherein the first viscosity level is lower than the second viscosity level to form a first layer at a porosity higher than that of the second layer.

6 . The method of claim 1 , wherein the first layer is formed from electrospinning polycaprolactone.

7 . The method of claim 1 , wherein the second layer is formed from spinning polycaprolactone/polyvinylpyrrolidone.

8 . The method of claim 1 , further comprising treating the epithelial support layer with at least one of a collagen or matrigel hydrogel material to alter a hydrogel stiffness as a function of depth away from the epithelial support layer.

9 . The method of claim 1 , wherein the first and second solutions are applied via positively charged nozzles, each at a predetermined rate and humidity environment and are attracted to a negative charge generated during the electrospinning.

10 . The method of claim 1 , wherein the electrospinning includes electrospinning directly on an epithelial support layer comprising track-etched polycarbonate membranes.

11 . A method for generating an electrospun mat for use in a three dimensional cell culture environment, comprising:
electrospinning a mat to facilitate intimate coupling between epithelial cell and a porous matrix, including co-spinning a first solution and a second solution, wherein the mat includes an epithelial support layer configured to intimately mate with epithelial cells cultured thereon.

12 . The method of claim 11 , further comprising integrating a hydrogel to the epithelial support layer to create a lower porosity surface for cell culture.

13 . The method of claim 11 , wherein the first and second solutions are applied via positively charged nozzles, each at a predetermined rate and humidity environment and are attracted to a negative charge generated during the electrospinning.

14 . The method of claim 13 , wherein a water-soluble polymer is formed with the porous matrix, and the method further comprises dissolving the water-soluble polymer in water.

15 . A method for generating a cell support interface for use in a three dimensional cell culture environment, comprising:
electrospinning a first layer using a first solution at a first viscosity level; and electrospinning a second layer using a second solution at a second viscosity level different from the first viscosity level, where the first layer and the second layer result in differing porosity levels for the first and second layers.

16 . The method of claim 15 , wherein the first viscosity level is higher than the second viscosity level to form the first layer at a porosity lower than that of the second layer.

17 . The method of claim 15 , wherein the first and second solutions are applied via positively charged nozzles, each at a predetermine rate and humidity environment and are attracted to a negative charge generated during the electrospinning.

18 . The method of claim 15 , where a water-soluble polymer is formed with the second layer and further comprising dissolving the water-soluble polymer.

19 . The method of claim 15 , wherein the first layer is formed from electrospinning polycaprolactone and wherein the second layer is formed from spinning polycaprolactone/polyvinylpyrrolidone.

20 . The method of claim 15 , further comprising treating the epithelial support layer with at least one of a collagen or matrigel hydrogel material to alter a hydrogel stiffness as a function of depth away from the epithelial support layer.

Assignees

• Charles Stark Draper Laboratory Inc

Inventors

• Christianson Rebecca
• Wiellette Elizabeth
• Sprachman Melissa
• Meehan-Qiu Alicica M
• Hapach Lauren A
• Hsi Peter
• Azizgolshani Hesham

Classifications

• C12M25/14 Primary

  Scaffolds; Matrices (in general C12N5/0068) · CPC title

• C12M25/02

  Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00) · CPC title

• D01D5/0007

  Electro-spinning (non-woven fabrics produced by electro-spinning D04H1/728) · CPC title

• C12M21/08

  for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00) · CPC title

• D10B2321/12

  polymers of cyclic compounds with one carbon-to-carbon double bond in the side chain · CPC title