Selective access to cryopreserved samples
Inventors
Childs, Richard W. • Cullis, Herb • Vasu, Sumithira • Broussard, Phillippe Jean • Clark, Kevin Douglas • Harting, Eric Kelsey
Assignees
Publication Number
US-9296500-B2
Publication Date
2016-03-29
Expiration Date
2030-05-04
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Methods and apparatus for selectively accessing a portion of a sterile cryopreserved sample are disclosed. The apparatus may include a container configured to receive the cryopreserved sample and having a first portion and a second portion, a heat sink chamber surrounding the first portion of the container, and a heat source adjacent to the second portion of the container. The chamber may be configured to maintain a non-accessed portion of the sample in a cryopreserved state. The heat source may be configured to separating an accessed portion of the sample from the non-accessed portion of the sample while maintaining the viability of the accessed portion while the non-accessed portion is maintained in the cryopreserved state.
Core Innovation
The invention provides methods and apparatus for selectively accessing a portion of a sterile cryopreserved sample. It includes a container holding the cryopreserved sample with distinct first and second portions, a heat sink chamber surrounding the first portion to maintain it in a cryopreserved state, and a heat source adjacent to the second portion. This system allows separation of an accessed portion from a non-accessed portion, maintaining viability of the accessed portion while preserving the remainder in a stably frozen state.
The problem addressed is the inability to selectively thaw or access a small portion of a frozen umbilical cord blood (UCB) unit or similar cryopreserved cell sample during the pre-transplant period without compromising the sterility, viability, or integrity of the remaining frozen portion. Existing methods require pre-freezing separation or do not allow selective thawing of small aliquots such as 1-2 ml. This limitation prevents early in vitro expansion of immune cells like NK cells or viral-reactive T cells prior to transplantation, which could reduce graft-versus-host disease (GVHD) and viral infection-related complications.
The disclosed embodiments include (1) an apparatus using a liquid nitrogen cooled block enclosure as a heat sink and heated severing bars that melt and fuse the plastic container to isolate and seal the accessed portion; (2) an apparatus employing coolant-filled bladders to maintain most of the sample frozen while a controlled heat source selectively thaws a small portion; (3) use of a patch layer that melts to seal tears occurring during severing; and (4) a release film layer placed between heating elements and the patch layer to prevent sticking and maintain container integrity during sealing. These technical arrangements enable removal or thawing of a small portion of the sample while preserving the remainder for later transplantation or use.
Claims Coverage
The patent includes multiple independent claims outlining methods and apparatus for selective access to cryopreserved samples, with several inventive features related to containers, heat sink chambers, heating and sealing elements, and sample types.
Selective access method using heat sink chamber and heat source
A method comprising placing a container with a cryopreserved sample in a heat sink chamber that surrounds a first portion, keeping a second portion outside the heat sink chamber. Creating a seal between first and second portions using a heat source and cutting along the seal to separate the sample into two portions.
Use of severing bars with heating elements
The heat source includes two severing bars, each having a heating element. Bars are tapered from a narrow leading edge to a wider trailing portion, providing localized heating and pressure to isolate and seal sample portions.
Temperature control of heating elements
Each heating element is covered by a heat-resistant, flexible cover and their temperature is controlled by a temperature controller, enabling precise melting of the container without damaging the sample.
Application of pressure for sealing
Pressing the severing bars against the container adjacent to the portion outside the heat sink chamber to apply mechanical force for melting and fusing opposing sides of the container to form a sterile seal.
Use of release film and patch layer
Placing a release film between the container and heat source with a higher melting point than the container prevents sticking to the heating elements. A patch layer made of plastic with melting temperature similar to the container is placed between the release film and container to fill tears during severing and ensure sealing integrity.
Cryopreserved sample types
The cryopreserved sample includes umbilical cord blood cells, lymphocytes, stem cells, red blood cells, white blood cells, modified cells, stromal cells, hybridoma cells, producer cells, pathogenic cells, epithelial cells, mesenchymal cells, sperm, embryos, biological cell parts, virus samples, rickettsial cells, vaccine materials, antigenic materials, cytokines, and hormones.
The claims cover methods and apparatus that enable localized, sterile partitioning of cryopreserved samples by combining a heat sink chamber, precisely heated and tapered sealing bars with temperature and pressure control, and additional sealing layers. These features collectively allow selective thawing or removal of sample portions while preserving the remaining frozen material in a viable, sterile condition.
Stated Advantages
Allows removal or thawing of a portion of a cryopreserved sample without compromising the integrity, sterility, or viability of the remaining portion.
Enables in vitro expansion of immune cells from a small aliquot prior to transplantation, potentially reducing graft-versus-host disease and viral infection-related mortality.
Minimizes thawing damage by producing a narrow melting channel and rapid refreezing due to tapered, thermally insulating severing bars.
Use of patch and release layers improves seal integrity and prevents tearing or contamination during severing.
Documented Applications
Selective thawing of small aliquots from frozen umbilical cord blood units to expand NK cells before transplantation to reduce graft-versus-host disease and enhance graft-versus-tumor effects.
Selective thawing of portions to expand virus-specific T cells ex vivo to reduce viral infection-related mortality post-transplant.
Selective access or thawing of portions from other frozen cell samples or biological material including frozen bone marrow, mobilized hematopoietic progenitor cells, plasma, and serum for therapeutic or research use.
Interested in licensing this patent?