System and method for in vitro blood vessel modeling

• US 9,595,206 B2
• Filed: 02/11/2009
• Issued: 03/14/2017
• Est. Priority Date: 02/11/2008
• Status: Active Grant

• Current Assignee: Charles Stark Draper Laboratory Incorporated
• Original Assignee: Charles Stark Draper Laboratory Incorporated, General Hospital Corporation
• Inventors: Hoganson, David M., Vacanti, Joseph P., Borenstein, Jeffrey T.
• Primary Examiner(s): Kim, Taeyoon
• Assistant Examiner(s): GOUGH, TIFFANY MAUREEN
• Application Number:
  US12/866,968
• Publication Number:
  US 20110053207A1
• Time in Patent Office: 2,953 Days
• Field of Search: 435/29
• US Class Current: 1/1
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: C08L 69/00 Compositions of polycarbonates; Compositions of derivatives of polycarbonates
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/34 Internal compartments or partitions
• CPC Class Codes: C12M 25/02 Membranes; Filters filters or filtration in general B01D24/00-B01D41/00
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture
• CPC Class Codes: G01N 33/5088 of vertebrates
• CPC Class Codes: G09B 23/28 for medicine

Abstract

The present invention provides an in vitro blood vessel model for investigation of drug induced vascular injury and other vascular pathologies. The in vitro blood vessel model provides two channels separated by a porous membrane that is coated on one side by an endothelial cell layer and is coated on the other side by a smooth muscle cell layer, wherein said model is susceptible to the extravasation of red blood cells across said porous membrane due to drug induced vascular injury.

Claims

1. An in vitro blood vessel model comprising two elongated channels separated by a porous membrane coated on one side by an endothelial cell layer and coated on the other side by a smooth muscle cell layer, wherein the model is susceptible to the extravasation of red blood cells across the porous membrane due to drug induced vascular injury, wherein one of the channels has:

a bottom formed by the porous membrane coated with the smooth muscle cell layer of the porous membrane;

opposing sidewalls extending from the bottom; and

a top that is open along a length of the channel.

2. The in vitro blood vessel model of claim 1 wherein the porous membrane has pore sizes ranging from approximately 0.1 to 30 μ

m and a porosity between approximately 5 and 50 percent.

3. The in vitro blood vessel model of claim 2 wherein said porous membrane has an 11 percent porosity, pore sizes ranging from 2.3 to 13 μ

m, and an average pore size of 6.4 μ

m.

4. The in vitro blood vessel model of claim 3 wherein said porous membrane is a track-etched polycarbonate membrane.

5. The in vitro blood vessel model of claim 4 wherein said porous membrane is coated with at least one of elastin, fibrin, fibronectin, laminin, hyaluronic acid, and collagen.

6. The in vitro blood vessel model of claim 1 wherein said two channels are a luminal channel and an open smooth muscle channel, wherein the open smooth muscle channel includes opposing beveled walls adjacent to the porous membrane.

7. The in vitro blood vessel model of claim 6 wherein at least one of the luminal channel and the smooth muscle channel includes at least one bifurcation.

8. The in vitro blood vessel model of claim 6 wherein said open smooth muscle channel is capped with a lid to seal the smooth muscle channel from an external atmosphere.

9. The in vitro blood vessel model of claim 1 wherein said two channels include offset inflow and outflow ponds that reduce non-physiological shear stress on the porous membrane.

10. The in vitro blood vessel model of claim 1 wherein fibroblasts are seeded on the smooth muscle cell layer to replicate an adventitial layer.

11. The in vitro blood vessel model of claim 1 wherein adipocytes are seeded on the smooth muscle cell layer to create perivascular fat tissue.

12. The in vitro blood vessel model of claim 6 wherein the porous membrane is remodeled by smooth muscle and endothelial cells over time to produce a membrane containing physiologic proteins that allows the extravasation under appropriate conditions.

13. An in vitro model of a small diameter blood vessel comprising:

a bottom layer defining a luminal channel;

a top layer defining an open top smooth muscle channel so that when the open top smooth muscle channel contains a fluid, the fluid is exposed to air;

a porous membrane sandwiched between the top layer and bottom layer to separate the channels, the porous membrane having;

a bottom side facing the luminal channel and being coated by an endothelial cell layer; and

a top side facing the open top smooth muscle channel and being coated by a smooth muscle cell layer,

wherein the porous membrane approximates an internal elastic lamina of blood vessels.

14. The model of claim 13, wherein the porous membrane is a track-etched polycarbonate membrane coated with at least one of elastin, fibronectin, fibrin, laminin, hyalauric acid, and collagen.

15. The model of claim 13, further comprising fibroblasts seeded above the smooth muscle cell layer.

16. The model of claim 13, wherein the open top smooth muscle channel is defined by opposing sidewalls in the top layer, enclosed on a bottom side by the porous membrane, open on a top side, and the opposing sidewalls are beveled towards the porous membrane.

17. The model of claim 13, wherein the bottom layer defines a first flow area pond in fluid communication with the luminal channel and the top layer defines a second flow area pond in fluid communication with the open top smooth muscle channel.

18. The model of claim 17, wherein:

the ponds are offset with respect to each other and the open top smooth muscle channel has a larger volume as compared to the luminal channel.

19. The model of claim 13, wherein the luminal channel is bifurcated to form an access port and further comprising a fitting coupled to the access port.

21. The model of claim 18, wherein the luminal channel and the open top smooth muscle channel are elongated and centrally aligned with the open top smooth muscle channel having a larger volume as compared to the luminal channel.

20. An in vitro model of a small diameter blood vessel comprising:

a bottom layer defining a luminal channel;

a top layer defining a smooth muscle channel so that when the smooth muscle channel contains a fluid, the fluid is exposed to air, wherein a volume of the smooth muscle channel is relatively larger than a volume of the luminal channel;

a porous membrane sandwiched between the top layer and bottom layer to separate the channels, the porous membrane having a bottom side facing the luminal channel and being coated by an endothelial cell layer and coated on a top side facing the smooth muscle channel by a smooth muscle cell layer, wherein the porous membrane approximates an internal elastic lamina of blood vessels,

wherein the smooth muscle channel has a bottom extending between opposing sidewalls and an open top.

22. The in vitro blood vessel model of claim 20 wherein the open top is open along an entire length of the smooth muscle channel.