Methods for inducing reversible stasis
First Claim
1. A method for cryopreserving biological material comprising:
- a) first incubating the biological material under anoxic conditions for an effective amount of time for the biological material to enter stasis; and
b) then cryopreserving the biological material.
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Abstract
The present invention concerns compositions and methods involving incubating biological materials under hypoxic or anoxic conditions to induce stasis or suspended animation. Methods of screening for compounds that induce stasis or compounds that increase the ability to undergo stasis are included. Such methods have ramifications for preserving biological materials as well as reducing or preventing trauma to biological materials. Also contemplated are methods for screening compounds that are active or more active under hypoxic conditions than normoxic conditions. Such methods can be used to identify antitumor compounds that would operate under hypoxic conditions in which tumor cells survive.
76 Citations
76 Claims
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1. A method for cryopreserving biological material comprising:
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a) first incubating the biological material under anoxic conditions for an effective amount of time for the biological material to enter stasis; and
b) then cryopreserving the biological material. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method for preserving biological material comprising:
a) incubating the biological material under hypoxic conditions for an effective amount of time for the biological material to enter stasis. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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29. A method for screening for an antitumor compound comprising:
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a) incubating a first anoxia-resistant organism under hypoxic conditions sufficient to permit the organism to enter stasis;
b) incubating the first organism with a candidate compound;
c) observing the first organism for viability; and
d) comparing the first organism'"'"'s viability against a second anoxia-resistant organism'"'"'s viability incubated under normoxic conditions in the presence of the candidate compound, wherein viability of the second organism and lack of viability of the first organism identifies the candidate compound as an anti-tumor compound. - View Dependent Claims (30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41)
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42. An antitumor composition comprising an antitumor compound identified by a process comprising:
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a) incubating a first anoxia-resistant organism under hypoxic conditions sufficient to induce stasis;
b) incubating the first organism with a candidate compound; and
c) comparing the first organism'"'"'s viability against a second anoxia-resistant organism'"'"'s viability incubated under normoxic conditions in the presence of the candidate compound, wherein the compound is an antitumor compound if the first anoxia-resistant organism is no longer viable and the second anoxia-resistant organism is viable after incubation with the candidate compound. - View Dependent Claims (43, 44, 45, 46, 47, 48, 49, 50, 51)
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52. A method for killing cancer cells in a patient with a tumor comprising administering to the patient an therapeutically effective amount of an antitumor compound identified by the process comprising:
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a) incubating a first anoxia-resistant organism under hypoxic conditions sufficient to permit the organism to enter stasis;
b) incubating the first organism with a candidate compound;
c) observing the first organism for viability; and
d) comparing the first organism'"'"'s viability against a second anoxia-resistant organism'"'"'s viability incubated under normoxic conditions in the presence of the candidate compound, wherein viability of the second organism and lack of viability of the first organism identifies the candidate compound as an anti-tumor compound. - View Dependent Claims (53, 54, 55, 56, 57)
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58. A method for screening for a compound that induces stasis in biological material comprising:
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a) incubating a first organism capable of stasis with a candidate compound; and
b) evaluating the first organism for stasis, wherein the compound is a stasis inducer if the first organism exhibits stasis after exposure to the compound. - View Dependent Claims (59, 60)
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61. A method of screening for a compound that improves the ability to undergo stasis comprising:
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a) incubating a first organism capable of undergoing stasis under hypoxic conditions;
b) exposing the first organism to a candidate compound;
c) incubating a second organism capable of undergoing stasis under the same hypoxic conditions as the first organism;
d) comparing the first organism and the second organism.
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62. A stasis inducer compound identified by a process comprising:
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a) incubating an organism capable of entering stasis with a candidate compound; and
b) evaluating the organism for stasis, wherein the compound is a stasis inducer if the organism exhibits stasis after incubation with the compound.
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63. A reversible stasis inducer compound identified by a process comprising:
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a) incubating an organism capable of entering stasis with a candidate compound;
b) comparing the organism with an organism capable of entering stasis that is no longer incubated with the candidate compound, wherein the compound is a stasis inducer if the organism exhibits stasis after incubation with the compound, but does not exhibit stasis when no longer incubated with the compound. - View Dependent Claims (64)
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- 65. A method of inducing stasis in biological material comprising administering to the biological material a stasis inducer compound.
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74. A method of inducing stasis in a cell of an organism comprising administering to the cell of the organism an effective amount of a stasis inducer compound identified by a process comprising:
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a) incubating an organism capable of entering stasis with a candidate compound;
b) evaluating the organism for loss of stasis after the compound is removed, wherein the compound is a stasis inducer if the organism exhibits stasis after incubation with the compound, but no longer exhibits stasis after the compound is removed.
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75. A method of inducing stasis in an organism comprising administering to the organism an effective amount of a stasis inducer compound identified by a process comprising:
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a) incubating an organism capable of entering stasis with a candidate compound;
b) evaluating the organism for loss of stasis after the compound is removed, wherein the compound is a stasis inducer if the organism exhibits stasis after incubation with the compound, but no longer exhibits stasis after the compound is removed.
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76. A method of identifying a modulator of stasis in a biological material undergoing stasis comprising:
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a) incubating a first biological material undergoing stasis with a candidate compound, wherein the first biological material is incubated under hypoxic conditions;
b) evaluating the first biological material for loss of stasis;
c) comparing the first biological material with a second biological material undergoing stasis but not incubated with the candidate compound, wherein a difference in stasis between the first and second biological materials identifies the compound as a candidate modulator of stasis.
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Specification