# US 9,518,898 B2 · Cryogenic storage container with sealing closure and methods of using the same

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- Number: US 9,518,898 B2
- Document ID: US9518898B2
- Status: Granted
- Kind: B2
- Application number: 14/077,707
- Filed: 2013-11-12
- Published: 2014-06-12
- Granted: 2016-12-13
- Earliest claimed: 2012-12-06
- Assignees: Cook Medical Technologies LLC
- Applicants: Cook Medical Technologies LLC
- Inventors: Jorge L. Jimenez-Rios
- Web page: https://patentgrep.com/patent/US9518898B2

## CPC

- G01N 1/28: Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00})
- A01N 1/147: Carriers for immersion in cryogenic fluid for slow freezing or vitrification
- C12M 45/22: {Means for packing or storing viable microorganisms (casings for storing cell samples A61B10/0096, preservation of living parts of the human or animal body A01N1/10)}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}

## Abstract

A container or vial for the cryopreservation and/or vitrification and storage of a biological sample, including cellular samples and embryos is described. The container comprises a sealable closure that isolates a biological sample held in the container lumen from the external environment and is suitable for storage of the sample in liquid nitrogen. Methods for introducing and sealing biological samples within the container for cryopreservation and/or vitrification and storage are also described.

## Claims

### Claim 1

A sealable container comprising: a body comprising a proximal open end and a sealed distal end and a variable diameter lumen extending there between, the lumen having an external surface and an internal surface, the internal surface tapering generally radially inwardly from the proximal open end to the distal end and forming a sealing portion and a sample receiving portion, a closure configured for removable attachment to the body wherein the closure comprises a capping member adapted for engagement with the proximal open end and a deformable member having a tapered external surface configured for sealing engagement with the sealing portion of lumen internal surface, at least two projections extending radially outwardly from the external surface of the lumen, the projections comprising a radially outward-most surface extending between the proximal open end and the sealed distal end of the body, wherein the outward-most surface has a substantially constant diameter.

### Claim 2 (depends on 1)

The container of claim 1 further comprising an intermediate space located between the external surface of the lumen and the outward-most surface of the projections.

### Claim 3 (depends on 1)

The container of claim 1 wherein the container body comprises a single unitary piece of injection molded material.

### Claim 4 (depends on 3)

The container of claim 3 wherein the material comprises polypropylene.

### Claim 5 (depends on 1)

The container of claim 1 wherein the closure and the body comprise the same material.

### Claim 6 (depends on 1)

The container of claim 1 wherein the closure and the body are constructed of a material that comprises the same or substantially similar thermal contraction properties.

### Claim 7 (depends on 1)

The container of claim 1 wherein the proximal open end of the body comprises a threaded portion and wherein the closure comprises threads adapted for mating engagement with the threaded portion of the container body.

### Claim 8 (depends on 1)

The container of claim 1 wherein the external surface of the deformable member comprises a male taper and the internal surface of the lumen comprises a female taper and wherein the male taper and female taper have corresponding taper angles such that an impermeable seal is established between the respective tapered surfaces.

### Claim 9 (depends on 1)

The container of claim 1 wherein the diameter of the internal surface of the lumen varies from about 2 mm to about 7 mm.

### Claim 10 (depends on 1)

The container of claim 1 wherein the diameter of the radially outwardly extending protrusions is approximately 12 mm.

### Claim 11 (depends on 1)

The container of claim 1 wherein the closure further comprises a stem extending therefrom and wherein the stem comprises a proximal end configured for holding a biological sample.

### Claim 12 (depends on 11)

The container of claim 11 wherein the stem comprises a proximal end that extends into the sample receiving portion of the internal lumen when the closure is secured to the container body.

### Claim 13

A method for preparing a biological sample for cryogenic storage comprising: a. obtaining sealable container comprising: i. a body comprising a proximal open end and a sealed distal end and a variable diameter lumen extending there between, the lumen having an external surface and an internal surface, the internal surface tapering generally radially inwardly from the proximal open end to the distal end and forming a sealing portion and a sample receiving portion, ii. a closure configured for removable attachment to the body wherein the closure comprises a capping member adapted for engagement with the proximal open end and a deformable member having a tapered external surface configured for sealing engagement with the sealing portion of lumen internal surface, iii. at least two projections extending radially outwardly from the external surface of the lumen, the projections comprising a radially outward-most surface extending between the proximal open end and the sealed distal end of the body, wherein the outward-most surface has a substantially constant diameter; b. introducing a biological sample into the lumen of the container; c. attaching the closure to the container body.

### Claim 14 (depends on 13)

The method of claim 13 wherein the biological sample is a suspension of cells, and the suspension is introduced into the distal end of the lumen.

### Claim 15 (depends on 13)

The method of claim 13 wherein the biological sample is an embryo suspended in a cryoprotectant.

### Claim 16 (depends on 15)

The method of claim 15 wherein the embryo is retained on a stem that extends from the capping member.

### Claim 17 (depends on 13)

The method of claim 13 further comprising submerging at least a portion of the container into a volume of liquid nitrogen.

### Claim 18 (depends on 13)

The method of claim 13 further comprising storing the container in a cryogenic freezing medium for a selected period of time.

### Claim 19

A cryogenic storage method comprising: a. introducing a biological sample into the lumen of a container, the container comprising: i. a body comprising a proximal open end and a sealed distal end and a variable diameter lumen extending there between, the lumen having an external surface and an internal surface, the internal surface tapering generally radially inwardly from the proximal open end to the distal end and forming a sealing portion and a sample receiving portion, ii. a closure configured for removable attachment to the body wherein the closure comprises a capping member adapted for engagement with the proximal open end and a deformable member having a tapered external surface configured for sealing engagement with the sealing portion of lumen internal surface, iii. at least two projections extending radially outwardly from the external surface of the lumen, the projections comprising a radially outward-most surface extending between the proximal open end and the sealed distal end of the body, wherein the outward-most surface has a substantially constant diameter; b. exposing at least a portion of the container to a cryogenic freezing medium.

### Claim 20 (depends on 19)

The method of claim 19 wherein the lumen of the container is pre-cooled to a temperature suitable for the vitrification of a biological sample prior to the step of introducing the biological sample into the lumen.