CRYOPRESERVATION METHOD AND DEVICE
First Claim
1. A device for the ultra-fast cooling and cryopreservation of living cells, the device comprising:
- a. a sample container having a base and cover that together contact and press a cell sample into a thin layer, whereby the cell sample is within 50 μ
m to 200 μ
m of a coolant; and
,b. a connection adapter connected to an OHP with an evaporator attached on one end to the OHP, and a condenser attached to the OHP opposite the evaporator, the adaptor designed and dimensioned for receiving the sample container, whereby the coolant is provided to the sample container.
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Accused Products
Abstract
A device and method suitable for the cryopreservation of all types of biological cells is described. In this method, an ultra-fast cooling/warming device system is used to achieve vitrification of individual cells or cell suspensions without cryoprotectant agents (CPA) or with a low concentration of CPAs (<1M), to attenuate the formation of intracellular ice crystal formation during cooling, and to minimize devitrification during subsequent warming. The device system applies oscillating heat pipe (OHP) and nanofluid techniques, and is built through microfabrication. Several devices may be networked to increase the total volume of cell samples that the cryopreservation system can process simultaneously.
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Citations
29 Claims
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1. A device for the ultra-fast cooling and cryopreservation of living cells, the device comprising:
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a. a sample container having a base and cover that together contact and press a cell sample into a thin layer, whereby the cell sample is within 50 μ
m to 200 μ
m of a coolant; and
,b. a connection adapter connected to an OHP with an evaporator attached on one end to the OHP, and a condenser attached to the OHP opposite the evaporator, the adaptor designed and dimensioned for receiving the sample container, whereby the coolant is provided to the sample container. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A network of two or more cryopreservation devices connected in parallel or in series, to process multiple cell sample volumes equal to between 1 ml and 20 ml.
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15. A device for the ultra-fast cooling and cryopreservation of living cells, the device comprising:
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a. at least one cell sample container constructed of a thermally conductive material, the container including a cell holding member with a cover whereby the cell holding member and cover are between 10 μ
m and 200 μ
m apart, the cover contacts the cell sample and spreads the cells into a thin block layer, the cell container also containing at least one interior coolant passage that directs the flow of coolant fluid past the cell sample at a distance of less than 200 μ
m;b. one or more connection adapters with a U-shaped design; and
,c. an OHP connected to fittings on the connection adapters and passing coolant fluid through a condenser on one end and through an evaporator on the opposite end. - View Dependent Claims (16, 17)
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18. A method for cell cryopreservation through direct vitrification of cell samples, comprising:
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a. pressing out a cell sample to a thickness of between 10 μ
m and 200 μ
m; and
,b. locating a cooling fluid proximate to the cell sample with the coolant fluid being within 200 μ
m of the cell sample, whereby heat transfer will occur at a rate of at least 106 K/min to vitrify the cell sample and thereby produce cryopreserved cells. - View Dependent Claims (19, 20, 21, 22, 23, 24)
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25. A method for warming cryopreserved cells, comprising:
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a. obtaining a vitrified cell sample in the form of a thin layer block with a thickness of between 10 and 200 μ
m; and
,b. placing the vitrified cell sample proximal to a flowing coolant, in a manner sufficient to cause heat transfer at a rate of at least 106 K/min, causing the cell sample to reach biological temperatures. - View Dependent Claims (26, 27, 28)
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29. A device for the cooling of living cells, the device comprising a sample container having a base and a cover, whereby the base receives the cells with the cover capable of being actuated to contact the cells and spread the cells into a single layer, with the cells located proximate to a coolant at a distance of between 50 μ
- m and 200 μ
m from the coolant, the base being made of thermal conductive material to allow for cooling of the cells at a rate of at least 106 K/min.
- m and 200 μ
Specification