Systems and methods for producing micro-engineered models of the human cervix

Patent metadata

Field	Value
Publication number	US-11920116-B2
Application number	US-202117352015-A
Country	US
Kind code	B2
Filing date	Jun 18, 2021
Priority date	Oct 22, 2015
Publication date	Mar 5, 2024
Grant date	Mar 5, 2024

Abstract

The presently disclosed subject matter provides systems and methods for producing a three-dimensional model of a human cervix. A microdevice is provided for culturing human cervical cells. The microdevice can include an upper microchannel including live ectocervical epithelial cells. The microdevice can include a lower microchannel including a first parallel lane and a second parallel lane including stromal media. The first and the second parallel lanes can be lined with live vascular endothelial cells. The lower microchannel can include a third parallel lane including uterine fibroblasts and live smooth muscle cells embedded in hydrogel. The first, second, and third lanes of the lower microchannel can be separated by protrusion structures. The third parallel lane can be positioned in the lower microchannel in between the first and the second parallel lanes. The microdevice can further include a porous membrane positioned in between the upper microchannel and the lower microchannel.

First claim

What is claimed is:

1. A microdevice for culturing human cervical cells, comprising:
an upper microchannel comprising live cervical epithelial cells;
a lower microchannel comprising:
a first parallel lane and a second parallel lane comprising stromal media, each of the first parallel lane and the second parallel lane being and wherein the first and the second parallel lanes are lined with live vascular endothelial cells,
a third parallel lane disposed between the first parallel lane and the second parallel lane and comprising a hydrogel comprising uterine fibroblasts and live smooth muscle cells embedded therein in hydrogel, wherein the first parallel, second parallel, and third parallel lanes of the lower microchannel are separated by
protrusion structures disposed between the first parallel lane and the third parallel lane and between the second parallel lane and the third parallel lane, a top surface of each of the protrusion structures being offset from a top surface of the lower microchannel wherein the third parallel lane is positioned in the lower microchannel in between the first and the second parallel lanes; and
a porous membrane positioned between the upper microchannel and the lower microchannel.

2. The microdevice of claim 1 , wherein the porous membrane is configured to pass nutrients from the upper microchannel to the lower microchannel.

3. The microdevice of claim 1 , wherein the upper microchannel and the lower microchannel have separate inlets through which biomaterials can be injected into the microdevice.

4. The microdevice of claim 1 , wherein the protrusion structures are configured to pin menisci of a hydrogel precursor solution injected into the third parallel lane of the lower microchannel and prevent the hydrogel precursor solution from spilling into the first parallel lane and the second parallel lane.

5. The microdevice of claim 1 , wherein the live cervical epithelial cells in the upper microchannel are perfused with epithelial media and wherein the live vascular endothelial cells in the lower microchannel are perfused with the stromal media in the first parallel lane and the second parallel lane.

6. A method of manufacturing a microdevice for culturing human cervical cells, comprising:
injecting hydrogel precursor solution comprising cervical stromal cells into the third parallel lane of the lower microchannel of the microdevice of claim 1 ;
injecting human cervical epithelial cells into the upper microchannel of the microdevice, resulting in an extracellular membrane coated surface of at least one epithelial monolayer;
perfusing the first parallel lane and the second parallel lane of the lower microchannel with stromal media; and
seeding vascular endothelial cells into the first parallel lane and the second parallel lane to form a lining of endothelial cells.

7. The method of claim 6 , wherein the hydrogel precursor solution injected into the third parallel lane is allowed to solidify into a hydrogel before injecting the human cervical epithelial cells into the upper microchannel.

8. A microdevice, comprising:
an upper microchannel;
a lower microchannel comprising:
a first parallel lane and a second parallel lane, each of the first parallel lane and the second parallel lane being lined with live vascular endothelial cells;
a third parallel lane disposed between the first parallel lane and the second parallel lane and comprising a hydrogel comprising cells therein; and
protrusion structures disposed between the first parallel lane and the third parallel lane and between the second parallel lane and the third parallel lane, a top surface of each of the protrusion structures being offset from a top surface of the lower microchannel; and
a porous membrane positioned between the upper microchannel and the lower microchannel.

9. The microdevice of claim 8 , wherein the porous membrane is configured to pass nutrients from the upper microchannel to the lower microchannel.

10. The microdevice of claim 8 , wherein the upper microchannel and the lower microchannel have separate inlets through which biomaterials can be injected into the microdevice.

11. The microdevice of claim 8 , wherein the protrusion structures are configured to pin menisci of a hydrogel precursor solution injected into the third parallel lane of the lower microchannel and prevent the hydrogel precursor solution from spilling into the first parallel lane and the second parallel lane.

12. The microdevice of claim 8 , wherein the upper microchannel comprises live cervical epithelial cells.

13. The microdevice of claim 8 , wherein the hydrogel comprises uterine fibroblasts and live smooth muscle cells embedded therein.

Assignees

• Univ Pennsylvania

Inventors

• Huh Dongeun
• Seo Jeongyun

Classifications

• C12M25/02 Primary

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

• C12M21/08 Primary

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

• C12M23/16

  Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093) · CPC title

• C12N5/0682

  Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells (oocytes C12N5/0609) · CPC title

• C12N2502/1347

  Smooth muscle cells · CPC title