Claims for Patent: 5,667,985

Summary for Patent: 5,667,985

Title:	Tissue biopsy cell suspender for cell analysis
Abstract:	A tissue biopsy cell suspender device includes a receptacle having a sealable opening. The receptacle contains a sufficient quantity of an aqueous medium to support an enzymatic dissociation of cells from a tissue biopsy sample. The device has a removable closure with a first sealing position and a second sealing position for sealing the opening in the receptacle. The device further includes a frangible container having a sufficient quantity of an enzyme therein to dissociate cells from the tissue biopsy sample. The frangible container is covered with a membrane that is permeable to water and to the enzyme. The frangible container is disposed within the receptacle so that the container is intact when the closure is in the first sealing position and the container is breached when the closure is in the second sealing position. When the frangible container is breached, the enzyme is released to permeate into the medium and for dissociation of the cells from the tissue sample.
Inventor(s):	O\'Leary; Robert K. (Lexington, MA), Stevens; Timothy (Warwick, NY), Griffith; Richard L. (Allendale, NJ)
Assignee:	Becton Dickinson and Company (Franklin Lakes, NJ)
Application Number:	08/718,850
Patent Claims:	1. A tissue biopsy cell suspender device comprising: a receptacle having a sealable opening, said receptacle containing a sufficient quantity of an aqueous medium suitable to support an enzymatic dissociation of cells from a tissue biopsy sample; a removable closure for sealing said opening, said closure having a first sealing position and a second sealing position; and a frangible container having a sufficient quantity of an enzyme therein to dissociate cells from the tissue biopsy sample, said frangible container being covered with a permeable membrane, said membrane being permeable to water and to said enzyme, said frangible container being disposed within said receptacle so that said container is intact when said closure is in said first sealing position, said container being breached when said closure is in said second sealing position, thereby to release said enzyme to permeate through said membrane into said medium. 2. The device of claim 1 wherein said receptacle is a tube having a closed end and an open end having a neck, said neck having at least one detent to engage said closure when said closure is positioned to seal said opening thereby defining said first sealing position. 3. The device of claim 2 wherein said closure is a resilient stopper comprising a socket having an inside surface with an inward shoulder thereon for receiving said neck of said tube, and wherein said detent comprises a groove on said tube positioned to releasably engage said shoulder. 4. The device of claim 2 wherein said socket further comprises a sufficient clearance to allow said closure to be moved onto said neck to beyond said detent to said second sealing position thereby breaching said frangible container. 5. The device of claim 1 wherein said medium is selected from the group consisting of aqueous minimum essential medium (M.E.M.) and balanced salts solution, (B.S.S.). 6. The device of claim 1 wherein said enzyme is selected from the group consisting of collagenases of the type I, type II, type III and type IV, ribonuclease, deoxribonuclease, amylase, hyaluronidase, elastase, papain, chymopapain, chymotrypsin, trpysin and mixtures thereof. 7. The device of claim 6 wherein said sufficient quantity of said enzyme comprises sufficient enzyme to provide a concentration of said enzyme in said medium between about 0.2 nanokatals to about 0.6 nanokatals. 8. The device of claim 1 wherein said frangible container is formed from glass, said enzyme being lyophilized and sealed in said container. 9. The device of claim 1 wherein said membrane is selected from the group consisting of decacetylated cellulose heat sealable film, polyethylene terephthalate film, polycarbonate film, polypropylene film. 10. The device of claim 9 wherein said membrane has a multiplicity of pores sized between about 0.1 microns and 10 microns with a pore density of between about 0.1.times.10.sup.6 to about 10.0.times.10.sup.6 per square centimeter. 11. The device of claim 10 wherein said membrane has a thickness between about 15 microns and 30 .mu.m. 12. The device of claim 11 wherein said membrane is formed from polyethylene terephthalate film with a thickness between about 20 microns and 30 microns and has between about 0.1.times.10.sup.6 and 10.times.10.sup.6 pores per square centimeter. 13. A tissue biopsy cell suspender device comprising: a tubular receptacle with a capacity between about one and five ml having a sealable opening, said receptacle containing between about 0.5 ml to about 1.5 ml of an aqueous minimum essential medium (M.E.M) to support an enzymatic dissociation of cells from a tissue biopsy sample; a removable closure for sealing said opening, said closure having a first sealing position and a second sealing position; and a frangible glass container having a sufficient quantity of an enzyme therein to dissociate cells from the tissue biopsy sample, said frangible container being covered with a permeable polyethylene terephthalate membrane, said membrane being permeable to water and to said enzyme, said frangible container being disposed within said receptacle so that said container is intact when said closure is in said first sealing position, said container being breached when said closure is in said second sealing position, thereby to release said enzyme to permeate through said membrane into said medium. 14. The device of claim 13 wherein said enzyme is collagenase in sufficient quantity to form a concentration of between about 43 nanokatals to about 51 nanokatals per milliliter in said medium. 15. The device of claim 13 wherein said enzyme is chymopapain in sufficient quantity to form a concentration of between about 0.22 nanokatals to about 0.44 nanokatals per milliter in said medium. 16. The device of claim 13 wherein said enzyme is a mixture of sufficient collagenase to form a concentration of between about 43 nanokatals to about 51 nanokatals per milliliter plus sufficient chymotrypsin to form a concentration of about 0.25 nanotakals per milliliter. 17. A method for releasing intact cells for analysis from a tissue biopsy sample comprising: providing a sample of tissue from a biopsy; removing a resealable closure from an opening of a tissue cell suspender device having a receptacle, said receptacle containing a sufficient quantity of an aqueous medium to support an enzymatic dissociation of cells from a tissue biopsy sample wherein said closure has a first sealing position and a second sealing position, said receptacle having a frangible container having a sufficient quantity of an enzyme therein to dissociate cells from the tissue biopsy sample, said frangible container being covered with a permeable membrane, said membrane being permeable to water and to said enzyme, said frangible container being disposed within said receptacle so that said container is intact when said closure is in said first sealing position, said container being breached when said closure is in said second sealing position, thereby to release said enzyme to permeate into said medium; introducing the tissue sample into said tissue cell suspender device; replacing said closure thereby sealing said receptacle; moving said closure to said second sealing position, thereby breaching said frangible container and releasing said enzyme into said medium; and incubating said tissue sample container for a sufficient time to allow said enzyme to permeate into said medium and dissociate the cells from said tissue.

Details for Patent 5,667,985

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Discure Medical, Llc	CHYMODIACTIN	chymopapain	For Injection	018663	11/10/1982		2039-03-29
Discure Medical, Llc	CHYMODIACTIN	chymopapain	For Injection	018663	08/21/1984		2039-03-29
Bausch & Lomb Incorporated	VITRASE	hyaluronidase	Injection	021640	05/05/2004		2039-03-29
Bausch & Lomb Incorporated	VITRASE	hyaluronidase	Injection	021640	12/02/2004		2039-03-29