Device and method for collecting and processing fat tissue and procuring microvessel endothelial cells to produce endothelial cell product

• US 5,372,945 A
• Filed: 07/31/1992
• Issued: 12/13/1994
• Est. Priority Date: 06/06/1985
• Status: Expired due to Term

• Current Assignee: Thomas Jefferson University (Main Line Health System)
• Original Assignee: Becton, Dickinson & Co
• Inventors: Williams, Stuart K., Prais, Alfred W., Jarrell, Bruce E., DiPisa, Joseph A. Jr., Alchas, Paul G.
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US07/923,384
• Time in Patent Office: 865 Days
• Field of Search: 435/1, 435/240.21, 435/262, 435/267, 435/271, 435/284, 435/286, 435/287, 435/288, 435/296, 435/311, 435/316, 435/803, 422/72, 422/101-103, 422/113, 210/632, 210/360.1, 210/380.1, 210/406, 210/472, 209/12, 209/19, 494/16, 494/27, 494/30, 494/36, 604/35, 604/190, 604/406, 128/749
• US Class Current: 435/267
• CPC Class Codes: A61F 2/062 Apparatus for the production of blood vessels made from natural tissue or with layers of living cells prostheses made from natural tissue or living cells A61L27/507
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3808 Endothelial cells
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: A61M 2202/0014 removed from the body
• CPC Class Codes: A61M 2202/0071 product to be retained or harvested, e.g. by pheresis
• CPC Class Codes: A61M 2202/08 Lipoids
• CPC Class Codes: A61M 2202/097 endothelial cells
• CPC Class Codes: A61M 2230/005 Parameter used as control input for the apparatus
• CPC Class Codes: C08L 89/06 derived from leather or skin , e.g. gelatin
• CPC Class Codes: C12M 45/09 by enzymatic treatment
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Methods and apparatus for collecting and processing tissue to produce an endothelial cell product having a vessel for rinsing, draining, digesting and isolating tissue. The vessel has a rinsing and digesting chamber for containing tissue during processing. An inlet in the rinsing and digesting chamber allows entry of rinsing solution and tissue from a liposuction device. A waste chamber in fluid communication with the rinsing and digesting chamber preferably connects with a vacuum source. An isolation chamber is separated from the rinsing and digesting chamber by a screen. An ampule in fluid communication with the isolation chamber includes a pair of ports controlled by valve devices to be selectively in fluid communication with the isolation chamber. After processing, the ampule isolates a pellet of endothelial cells and the valve devices permit the pellet to be in fluid communication with the ports. The method includes providing the vessel, introducing tissue to be processed, orienting the vessel to screen the tissue, introducing an enzyme and agitating to digest the tissue, centrifuging the vessel to transfer the cells from the digested tissue, and isolating the cells for retrieval.

Claims

1. A method for collecting and processing tissue for producing an endothelial cell product comprising:

providing a collecting vessel for processing tissue to produce an endothelial cell product, the vessel having a rinsing and digesting chamber in fluid communication with a separate waste chamber and an isolation chamber connected to the rinsing and digesting chamber, wherein a screen separates at least the rinsing and digesting chamber and the waste chamber;

introducing tissue to be processed into the rinsing and digesting chamber;

introducing rinsing solution into the rinsing and digesting chamber;

orienting the vessel to screen the tissue to be processed of rinsing solution passed into the waste chamber;

introducing an enzyme into the rinsing and digesting chamber;

heating the tissue and the enzyme for a sufficient time and temperature while agitating the rinsing and digesting chamber to digest the tissue with the enzyme;

centrifuging the vessel to transfer cells from the digested tissue from the rinsing and digesting chamber into the isolation chamber; and

isolating the cells as a pellet of microvessel endothelial cells.

2. The method of claim 1 with the further step of isolating the cells from the digested tissue in an ampule as a pellet of microvessel endothelial cells by application of a valve means selectively connecting the isolation chamber and the ampule.

3. A device for digesting and processing tissue to produce an endothelial cell product comprising:

a single vessel comprising three chambers;

a digestion chamber;

a separate waste chamber; and

a separate isolation chamber, said waste and isolation chambers in fluid communication with the digestion chamber, wherein the isolation chamber is selectively connected by a first valve to an ampule for receiving said endothelial cell product, and wherein the digestion chamber is selectively and alternately in fluid communication with the waste chamber and the isolation chamber,

whereby the device allows the digestion of tissue by connecting the digestion chamber to the waste chamber, and processing of tissue by connecting the digestion chamber to the isolation chamber to produce endothelial cell product within said single vessel under sterile conditions.

4. The device of claim 3 further comprising a normally closed check valve connecting the digestion chamber and the waste chamber.

5. The device of claim 4 further comprising an internal vent tube for permitting flow through the check valve, the vent tube extending from the waste chamber into the isolation chamber and a valve regulating fluid flow between the waste chamber and the isolation chamber.

6. The device of claim 3 further comprising a second valve, each of said first and second valves having at least two positions, wherein in its first position the first valve allows communication between the isolation chamber and the ampule, and when in its first position the second valve allows communication between the ampule and an outlet port, and wherein, in a second position the valves are closed to isolate endothelial cell product within the ampule.

7. The device of claim 3 further comprising at least one inlet port affixed to the vessel in communication with the digestion chamber for introducing collected fat, rinse and enzyme solutions into said digestion chamber.

8. A device for collecting and processing tissue to produce an endothelial cell product, comprising a vessel for rinsing, draining, digesting and isolating tissue the vessel comprising:

a rinsing and digesting chamber in the vessel for collecting and processing tissue;

an inlet connected to the rinsing and digesting chamber for receiving rinsing solution and also for receiving tissue from a liposuction means for withdrawing tissue from a subject;

a waste chamber in fluid communication with the rinsing and digesting chamber, the waste chamber further comprising a connection for fluid communication with a vacuum source;

a first screen separating the rinsing and digesting chamber from the waste chamber wherein waste fluid comprising rinsing solution flows from the rinsing and digesting chamber through the first screen to the waste chamber;

an isolation chamber in fluid communication with and separated from the rinsing and digesting chamber by a second screen to allow cells from .rinsed, digested and processed tissue produced under sterile conditions to be isolated;

an ampule connected to and in selective fluid communication with said isolation chamber;

a pair of ports connected to the ampule, one of the ports providing a selective connection between the isolation chamber and the ampule and another of the ports providing selective communication between the ampule and an external environment; and

valve means operatively associated with each of the ports to selectively control fluid communication between each of the ports, the ampule, the external environment and the isolation chamber,

whereby endothelial cells disposed in the ampule are isolated by selective operation of the valve means.

9. A device for digesting and processing tissue to produce an endothelial cell product comprising:

a single vessel comprising four chambers;

(a) an enzyme storage chamber;

(b) a separate waste chamber;

(c) a separate digestion chamber alternately and selectively connected to the waste chamber and the enzyme chamber; and

(d) a separate isolation chamber selectively connected to the digestion chamber, wherein the isolation chamber is selectively connected by a first valve to an ampule for receiving said endothelial cell product, whereby the device allows the digestion of tissue by connecting the digestion chamber to the enzyme storage chamber and the waste chamber and processing of tissue by connecting the digestion chamber to the isolation chamber to produce endothelial cell product within a single vessel under sterile conditions.

10. The device of claim 9 wherein said enzyme storage chamber selectively communicates with said digestion chamber through a valve and said enzyme storage chamber communicates with an external environment through a port.

11. The device of claim 10 wherein the valve between said enzyme storage chamber and said digestion chamber is a normally closed check valve which opens when pressure increases inside said enzyme storage chamber upon centrifugation of said device.

12. The device of claim 9 wherein said digestion chamber further comprises a means to introduce tissue to be digested within said digestion chamber, and a means to introduce rinsing fluid into said digestion chamber for rinsing said tissue.