Apparatus for growing tissue specimens in vitro

• US 5,153,136 A
• Filed: 01/30/1990
• Issued: 10/06/1992
• Est. Priority Date: 07/22/1988
• Status: Expired due to Term

• Current Assignee: Miriam Hospital (Lifespan Corporation)
• Original Assignee: Herman H. Vandenburgh
• Inventors: Vandenburgh, Herman H.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Beisner, William H.
• Application Number:
  US07/472,347
• Time in Patent Office: 980 Days
• Field of Search: 435/1, 435/240.23, 435/240.241, 435/240.243, 435/284-286, 435/287, 435/289, 435/299-301, 435/316, 435/809, 435/3, 73/794, 73/806, 73/811
• US Class Current: 435/286.1
• CPC Class Codes: C12M 25/04 in combination with well or multiwell plates, i.e. culture inserts
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture

Abstract

An apparatus for growing tissue specimens in vitro includes an expandable membrane for receiving a tissue specimen thereon, a mechanism for expanding the membrane and the tissue specimen, and a controller for controlling the expanding mechanism. The controller is operative for applying an activity pattern to the membrane and a tissue specimen thereon which includes simultaneous continuous stretch activity and repetitive stretch and release activity. The continuous stretch activity and the repetitive stretch and release activity simulate the types of activity to which cells are exposed in vivo due to growth and movement, respectively, and they cause the cells of tissue speciments grown in the apparatus to develop as three-dimensional structures similar to those grown in vivo.

Claims

1. An apparatus for growing a tissue specimen in vitro comprising:

(a) expandable support means for supporting the tissue specimen and constructed so as to permit attachment of the tissue specimen thereto, said support means being expandable to effect stretching of the tissue specimen;

(b) means for mechanically expanding said support means to stretch the tissue specimen; and

(c) control means for controlling said mechanical expanding means to both gradually and substantially continuously expand said expandable support means over a prolonged period of time and to simultaneously repetitively expand and release said expandable support means over said period of time.

2. The apparatus of claim 1, wherein said control means controls said mechanical expanding means so as to expand and release said expandable support means by between 0.02% and 60% during said expand and release activity.

3. The apparatus of claim 1, wherein said control means controls said mechanical expanding means so as to periodically repetitively stretch and release the tissue specimen during activity periods of at least approximately five seconds duration.

4. In the apparatus of claim 3, said activity periods being separated by rest periods of at least approximately five seconds duration during which the tissue specimen is subjected to said gradual and substantially continuous stretch activity but not subjected to significant stretch and release activity.

5. In the apparatus of claim 1, said control means controlling said mechanical expanding means to gradually and substantially continuously stretch the tissue specimen at a rate of between 0.005 mm/hr and 1.0 mm/hr.

6. In the apparatus of claim 1, said support means comprising a substantially circular resiliently flexible membrane having an upwardly facing side and a downwardly facing side, said support means being operative for supporting the tissue specimen on the upwardly facing side of said membrane, said expanding means engaging the downwardly facing side of said membrane to deform said membrane and to thereby stretch the tissue specimen.

7. In the apparatus of claim 6, said support means further comprising a substantially circular support ring on said membrane extending upwardly from the upwardly facing side thereof, said support ring having an inwardly facing attachment surface thereon which has a sufficiently roughened texture to permit attachment of the tissue specimen thereto.

8. In the apparatus of claim 7, said support ring comprising a ring of screening which substantially defines the inwardly facing attachment surface thereof, said ring of screening being made of a biocompatible material.

9. In the apparatus of claim 8, said ring of screening being made of stainless steel.

10. In the apparatus of claim 8, said ring of screening having between 4×

4 and 250×

250 meshes/in, having a wire diameter of between 0.04 mm and 3.76 mm, and a mesh opening width of between 0.06 mm and 2.6 mm.

11. In the apparatus of claim 1, said support means comprising a flexible membrane having opposite first and second ends and having an upwardly facing side and a downwardly facing side, said mechanical expanding means being operative for stretching said membrane by separating said first and second ends.

12. In the apparatus of claim 11, said support means further comprising first and second support walls extending upwardly from the upwardly facing side of said membrane, said first and second support walls having first and second substantially opposed attachment surfaces thereon, respectively, which have sufficiently roughened textures to permit attachment of the tissue specimen thereto, said support walls being moved apart by said expanding means with said first and second ends and being disposed in substantially perpendicular relation to the direction of separating movement when said first and second ends are separated.

13. In the apparatus of claim 12, said first and second support walls comprising screening elements which substantially define said attachment surfaces thereon, said screening elements being made of a biocompatible material.

14. In the apparatus of claim 13, said screening elements being made of stainless steel.

15. In the apparatus of claim 14, said screening elements having between 4×

4 and 250×

250 meshes/in, having a wire diameter of between 0.04 mm and 3.76 mm, and a mesh opening width of between 0.06 mm and 2.6 mm.