Pulsatile Perfusion Bioreactor for Mimicking, Controlling, and Optimizing Blood Vessel Mechanics

Patent metadata

Field	Value
Publication number	US-2016298073-A1
Application number	US-201615071354-A
Country	US
Kind code	A1
Filing date	Mar 16, 2016
Priority date	Apr 7, 2015
Publication date	Oct 13, 2016
Grant date	—

Abstract

A pulsatile perfusion bioreactor for culturing one or more engineered blood vessels having a lumen and a wall is provided. The bioreactor includes a chamber for holding the engineered blood vessel and cell culture media; a mechanical property monitoring system for measuring axial tensile stress and strain, circumferential tensile stress and strain, and/or shear stress imparted on the vessel wall; and a pump system for delivering cell culture media through the vessel lumen, wherein the vessel is exposed to a composite pressure waveform and a composite flow waveform as the media is delivered there through. The pump system includes a steady flow and peristaltic pumps. Further, the composite pressure and flow waveforms each include a mean component, a fundamental frequency component, and a second harmonic frequency component. The bioreactor also includes a computer interface for monitoring and adjusting the composite waveforms to maintain a predetermined stress levels.

First claim

What is claimed:

1 . A pulsatile perfusion bioreactor for culturing one or more engineered blood vessels having a lumen and a wall, the pulsatile perfusion bioreactor comprising:
a chamber for holding the engineered blood vessel and cell culture media; a mechanical property monitoring system for measuring and controlling axial tensile stress, circumferential tensile stress, shear stress, or a combination thereof imparted on the wall of the engineered blood vessel and for measuring and controlling axial stretch, circumferential stretch, or a combination thereof imparted on the wall of the engineered blood vessel; a pump system for delivering cell culture media through the lumen of the engineered blood vessel, wherein the engineered blood vessel is exposed to a composite pressure waveform and a composite flow waveform as the cell culture media is delivered through the lumen, the pump system comprising a steady flow pump and a peristaltic pump, wherein the composite pressure waveform comprises a mean pressure component, a first harmonic frequency pressure component, and a second harmonic frequency pressure component, and wherein the composite flow waveform component comprises a mean flow component, a first harmonic frequency flow component, and a second harmonic frequency flow component; and a computer interface for monitoring and adjusting the composite pressure waveform, the composite flow waveform, or a combination thereof to maintain a predetermined axial tensile stress level, a predetermined circumferential stress level, a predetermined shear stress level, or a combination thereof.

2 . The bioreactor as in claim 1 , wherein the composite pressure waveform and the composite flow waveform are derived from a pressure waveform and a flow waveform of a native blood vessel, wherein the engineered blood vessel is a replacement for the native blood vessel.

3 . The bioreactor as in claim 1 , wherein the steady flow pump delivers the mean pressure component of the composite pressure waveform and the mean flow component of the composite flow waveform.

4 . The bioreactor as in claim 1 , wherein the peristaltic pump delivers a pulsatile flow of cell culture media through the lumen, wherein the peristaltic pump comprises a first pump head and a second pump head, wherein the first pump head provides the first harmonic frequency pressure component of the composite pressure waveform and the first harmonic frequency flow component of the composite flow waveform, and wherein the second pump head provides the second harmonic frequency pressure component of the composite pressure waveform and the second harmonic frequency flow component of the composite flow waveform.

5 . The bioreactor as in claim 4 , wherein the peristaltic pump further comprises a third pump head, wherein the third pump head provides a third harmonic frequency pressure component of the composite pressure waveform and a third harmonic frequency flow component of the composite flow waveform.

6 . The bioreactor as in claim 1 , further comprising a compliance chamber.

7 . The bioreactor as in claim 1 , further comprising a pressure transducer.

8 . The bioreactor as in claim 1 , further comprising a stepper motor controlled pinch valve.

9 . The bioreactor as in claim 1 , further comprising a camera for measuring the engineered blood vessel length, wall diameter, and wall thickness.

10 . The bioreactor as in claim 1 , wherein the chamber is located in an incubator.

11 . The bioreactor as in claim 1 , wherein the engineered blood vessel comprises a natural material or a synthetic material.

12 . The bioreactor as in claim 1 , wherein the engineered blood vessel includes endothelial cells.

13 . The bioreactor as in claim 1 , wherein the engineered blood vessel includes smooth muscle cells.

14 . A method of culturing a one or more engineered blood vessels having a lumen and a wall inside a pulsatile perfusion bioreactor, the method comprising:
inserting the engineered blood vessel to be cultured into a chamber; filling the chamber with cell culture media; delivering cell culture media through the lumen of the engineered blood vessel via a pump system, wherein the engineered blood vessel is exposed to a composite pressure waveform and a composite flow waveform as the cell culture media is delivered through the lumen, the pump system comprising a steady flow pump and a peristaltic pump, wherein the composite pressure waveform comprises a mean pressure component, a first harmonic frequency pressure component, and a second harmonic frequency pressure component, and wherein the composite flow waveform component comprises a mean flow component, a first harmonic frequency flow component, and a second harmonic frequency flow component; measuring axial tensile stress, circumferential tensile stress, shear stress, axial stretch, circumferential stretch, or a combination thereof imparted on the wall of the engineered blood vessel via a mechanical property monitoring system; monitoring and adjusting the composite pressure waveform, the composite flow waveform, or a combination thereof to maintain a predetermined axial tensile stress level, a predetermined circumferential stress level, a predetermined shear stress level, a predetermined axial stretch level, a predetermined circumferential stretch level, or a combination thereof via a computer interface.

15 . The method as in claim 14 , wherein the composite pressure waveform and the composite flow waveform are derived from a pressure waveform and a flow waveform of a native blood vessel, wherein the engineered blood vessel is a replacement for the native blood vessel.

16 . The method as in claim 14 , wherein the steady flow pump delivers the mean pressure component of the composite pressure waveform and the mean flow component of the composite flow waveform.

17 . The method as in claim 14 , wherein the peristaltic pump delivers a pulsatile flow of cell culture media through the lumen, wherein the peristaltic pump comprises a first pump head and a second pump head, wherein the first pump head provides the first harmonic frequency pressure component of the composite pressure waveform and the first harmonic frequency flow component of the composite flow waveform, and wherein the second pump head provides the second harmonic frequency pressure component of the composite pressure waveform and the second harmonic frequency flow component of the composite flow waveform.

18 . The method as in any of claim 17 , wherein the peristaltic pump further comprises a third pump head, wherein the third pump head provides a third harmonic frequency pressure component of the composite pressure waveform and a third harmonic frequency flow component of the composite flow waveform.

19 . The method as in claim 15 , wherein the pulsatile perfusion bioreactor includes a compliance chamber, wherein the compliance chamber facilitates adjustment of the composite pressure waveform.

20 . The method as in claim 15 , wherein pressure is measured via a pressure transducer and a stepper motor controlled pinch valve is utilized to adjust resistance within the pulsatile perfusion bioreactor, wherein adjusting the resistance results in an adjustment to the pressure.

Assignees

• Univ South Carolina

Inventors

• Eberth John F
• Shazly Tarek
• Zhou Boran
• Prim David A
• Gore Conrad Michael

Classifications

• C12N5/0691

  Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels · CPC title

• C12M29/12 Primary

  Pulsatile flow · CPC title

• C12M35/04

  Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli · CPC title

• C12M29/10

  Perfusion · CPC title

• C12M41/40

  of pressure · CPC title