Vascularized in vitro perfusion devices, methods of fabricating, and applications thereof

• US 10,392,595 B2
• Filed: 07/06/2016
• Issued: 08/27/2019
• Est. Priority Date: 07/06/2015
• Status: Active Grant

• Current Assignee: Advanced Solutions Life Sciences, LLC
• Original Assignee: Advanced Solutions Life Sciences, LLC
• Inventors: Golway, Michael W., Hoying, James B.
• Primary Examiner(s): Davis, Ruth A
• Application Number:
  US15/202,675
• Publication Number:
  US 20170009194A1
• Time in Patent Office: 1,147 Days
• CPC Class Codes: C12M 23/16 Microfluidic devices; Capillary tubes integrated microfluidic structures B01L3/5027; microreactors B01J19/0093
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 33/00 Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00
• CPC Class Codes: C12N 2513/00 3D culture
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0691 Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5064 Endothelial cells

Abstract

A bona fide adaptable in vitro microcirculation model is provided by integrating a 3-D printed network of endothelial-cell lined perfusion channels, formed via sacrificial casting in a gel matrix, with a native, adaptable microvasculature matured from native microvessels added to the gel matrix. Responsive vascular adaptation exhibited by the in vitro microcirculation is physiologically relevant. Methods for fabricating, devices, models and investigative platforms for pharmaceutical applications, vascular mechanism and microvessel-parenchyma interaction studies, and vascularizing strategies for tissue engineering applications are also disclosed.

Claims

1. A vascularized in vitro perfusion device comprising an adaptable microcirculation, said device comprising:

a supporting structure comprising a gel matrix,

a fabricated network of microfluidic endothelial cell-lined channels, said fabricated network comprising an inlet channel, an outlet channel, and at least one cross channel connecting the inlet channel to the outlet channel, said cross channel positioned at least partially within the gel matrix,

an inlet port in fluid communication with the fabricated network, and an outlet port in fluid communication with the fabricated network, and

a neovasculature, said neovasculature being derived from intact native microvessels incorporated into the gel matrix and subject to maturing conditions comprising endothelial sprouting and an inosculation between at least two endothelial sprouts respectively derived from the at least one cross channel of the fabricated network and the neovasculature to form a lumen connection therebetween, said neovasculature thus in lumen continuity with said at least one cross channel through the inosculation,

wherein the fabricated network is in vascular communication and lumen continuity with the neovasculature to form an adaptable microcirculation.

2. The device according to claim 1, further comprising an enclosure comprising a housing.

3. The device according to claim 2, wherein the housing comprises a biocompatible plastic polymer selected from acrylic, styrene and carbonate polymers.

4. The device according to claim 2, wherein the at least one inlet port and at least one outlet port are accessed exterior to the enclosure.

5. The device according to claim 1, wherein the gel matrix is selected from a collagen gel, a fibrin gel, and combinations thereof.

6. The device according to claim 1, wherein the intact native microvessels are derived from adipose tissue.

7. The device according to claim 1, wherein the vascular communication is formed from perfusion-driven sprouting from the fabricated network of endothelial cell-lined channels, and inosculation between at least two sprouts and the neovasculature.

8. The device according to claim 1, wherein the inlet channel and the at least one cross channel have a cross-sectional diameter ratio between about 1.1:

1 and about 5;

1.

9. The device according to claim 8, comprising at least two cross channels having different cross-sectional diameters such that a hemodynamic gradient is formed between the cross channels upon perfusion.

10. The device according to claim 1, further comprising an external perfusion control system in operational communication with the fabricated network through the inlet port.

11. The device according to claim 1, wherein adaptable comprises an ability to undergo vascular differentiation and/or positive vascular remodeling in response to at least one perfusion-driven hemodynamic force or stimuli.

12. The device according to claim 11, wherein the at least one hemodynamic force is one or more of intravascular pressure and shear stress.

13. The device according to claim 1, further comprising living cells, said cells populating the gel matrix and being derived from one or more tissue types.

14. The device according to claim 13, wherein the cells are all from the same tissue type.

15. The device according to claim 13, wherein the gel matrix comprises at least two sections and each section is populated with cells from a distinct tissue type.

16. The device according to claim 13, wherein the living cells comprise human cells.

17. The device according to claim 13, wherein the living cells are selected from one or more of normal cells, diseased cells, stem cells, endothelial cells, stromal cells, epithelial cells, neuronal cells, connective cells, myocardial cells, hepatocytes, renal cells, tumor cells, liver cells, pancreatic cells, muscle cells, brain cells, kidney cells, and patient-specific cells.

18. The device according to claim 13, wherein adaptable comprises an ability to undergo vascular remodeling in response to at least one cue provided by the living cells.

19. The device according to claim 18, wherein the at least one cue is provided by the living cells in response to an agent delivered by perfusion.

20. The device according to claim 1, wherein the vascular communication is formed from one of perfusion-driven sprouting, cell-driven sprouting, growth factor-driven sprouting, and matrix driven-sprouting from the fabricated network of endothelial cell-lined channels, and inosculation between at least two sprouts and the neovasculature.