AUTOMATED, MULTIFUNCTIONAL, ENGINEERED CARDIAC TISSUE CULTURE AND TESTING BIOREACTOR SYSTEM

• US 20170260488A1
• Filed: 08/25/2015
• Published: 09/14/2017
• Est. Priority Date: 09/05/2014
• Status: Active Grant

• Current Assignee: Icahn School of Medicine at Mount Sinai
• Original Assignee: Icahn School of Medicine at Mount Sinai
• Inventors: Costa, Kevin D., Backeris, Peter
• Granted Patent: US 10,119,109 B2
• Time in Patent Office: Days
• 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 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/22 Transparent or translucent parts glassware for laboratory use B01L3/00
• CPC Class Codes: C12M 23/38 Caps; Covers; Plugs; Pouring means
• CPC Class Codes: C12M 23/50 Means for positioning or orientating the apparatus C12M41/08 takes precedence
• CPC Class Codes: C12M 31/10 by light emitting elements located inside the reactor, e.g. LED or OLED
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00

Abstract

An improved tissue engineering bioreactor and testing platform has been designed that integrates multiple testing and stimulation capabilities. The system allows for growth of multiple tissue strips in parallel with mechanical and electrical stimulation, media perfusion, and the automated monitoring of contractile force and extracellular electrical activity. The system is designed to be low-cost and scalable, to provide for high-content, biofidelic, non-destructive testing of engineered muscle tissue performance that is conventionally measured using muscle-bath systems.

Claims

1. A bioreactor system that is configured to provide length control of engineered tissue and to measure one or more properties of the engineered tissue comprising:

a housing that includes a culture well;

at least one first flexible post suspended in the culture well;

at least one second rigid post suspended in the culture well, the second post being spaced from the first post such that the engineered tissue is grown therebetween;

a first device for measuring a contractile force of the engineered tissue, the device including the at least one first post which comprises a fiber-optic that is operatively coupled to a light source and configured to flex when the engineered tissue contracts, the first device further including a sensor for detecting a degree of movement of the at least one first post; and

a second device for controlling the length of the engineered tissue, the second device including the at least one rigid second post and an actuator that is operatively coupled to the at least one second post for moving the at least one second post in a controlled manner within the culture well so as to cause a change in a distance between the at least one first post and the at least one second post and thereby control the length of the engineered tissue.

2. The bioreactor system of claim 1, wherein the housing includes a base which contains the culture well and a removable lid with the second device being fixedly attached to the lid.

3. The bioreactor system of claim 1, wherein three or more first posts and three or more second posts are suspended in the culture well for growing three or more engineered tissue specimens.

4. The bioreactor system of claim 3, wherein the three or more first posts are arranged linearly and the three or more second posts are arranged linearly so as to define at least sets of facing posts, each set of posts supporting one engineered tissue specimen resulting in three or more engineered tissue specimens being grown in parallel.

5. The bioreactor system of claim 1, wherein the first device includes a base which is coupled to the housing to position and suspend the at least one first post in the culture well.

6. The bioreactor system of claim 1, wherein the first device includes a fiber optic LED emitter that is coupled to the fiber-optic (first post) for generating a light spot that contacts the sensor which comprises a position-sensitive detector which is configured to measure tissue contractions by measuring the degree of movement of the fiber-optic by measuring a position of the light spot.

7. The bioreactor system of claim 6, wherein a floor of the culture well is formed of a material that allows passage of light from the fiber-optic to the position-sensitive detector which is disposed below the culture well.

8. The bioreactor system of claim 1, wherein the fiber-optic has a diameter of about 0.55 mm.

9. The bioreactor system of claim 2, wherein the fiber-optic passes through the lid with an upper end of the fiber-optic proximate the lid representing a more rigid section of the first post, with a lower section thereof representing a flexible section that flexes in response to applied forces due to tissue contractions.

10. The bioreactor system of claim 1, wherein the second post comprises a rigid tungsten post.

11. The bioreactor system of claim 1, wherein the actuator comprises a linear actuator that is operatively coupled to a post holder to which the at least one second post is coupled.

12. The bioreactor system of claim 11, wherein the post holder is operatively coupled to linear slides that under actuation of the linear actuator results in linear movement of the post holder which results in linear movement of the engineered tissue due to one end of the engineered tissue being coupled to the second post.

13. The bioreactor system of claim 1, further including a pair of electrodes disposed within the culture well outside of the at least one first post and the at least one second post for supplying electrical stimulation across a length of the engineered tissue.

14. The bioreactor system of claim 13, wherein the electrodes comprise graphite sheet electrodes arranged in the culture well such that the at least one first post and the at least one second post are disposed therebetween.

15. The bioreactor system of claim 1, wherein each of the first and second devices is operatively coupled to a controller for controlling operation and functionality thereof.

16. The bioreactor system of claim 1, further including a mechanism for measuring action potentials generated by the engineered tissue during contractions.

17. The bioreactor system of claim 16, wherein the mechanism includes an amplifier that is configured such that inputs thereof are connected to the at least one second post enabling extracellular recording of action potentials generated by the engineered tissue.

18. The bioreactor system of claim 1, wherein the culture well includes built-in fluid exchange ports that allow for the perfusion of media and testing agents into and out of the culture well.

19. The bioreactor system of claim 2, further including a pair of stimulating electrodes that are detachably attached to the lid and positioned in the culture well such that the at least one first post and the at least one second post are disposed between the electrodes.

20. The bioreactor system of claim 19, wherein the electrodes include conductive magnets for fixedly, yet detachably, attaching the electrodes to the lid such that the electrodes are maintained in fixed positions during use, are easily removable from the lid for cleaning and are quickly replaceable after cleaning thereof.