Automated system for cryopreservation of oocytes, embryos, or blastocysts

Abstract

An automated system and method of cryopreservation and reanimation of oocytes, embryos, or blastocysts is disclosed. One or more oocytes or embryos are positioned in a processing container, the processing container being configured to allow fluid to flow into and out of the processing container, where two or more fluids flow into and out of the processing container with oocytes or embryos therein. The temperature of the fluid may be controlled in the processing container according to predetermined requirements. The flowing of the fluids may be controlled by a central controller adapted to control one or more valves.

1 . A method of cryopreserving or reanimating oocytes, embryos or blastocysts, comprising:
 positioning one or more oocytes, embryos or blastocysts in a processing container, the processing container configured to allow two or more fluids to flow into and out of the processing container;   flowing the two or more fluids into and out of the processing container with the one or more oocytes, embryos or blastocysts therein.   
     
     
         2 . The method of  claim 1 , further comprising:
 controlling one or more characteristics of at least one of the two or more fluids flowing into and out of the processing container according to one or more process parameters.   
     
     
         3 . The method of  claim 2 , wherein the one or more of characteristics comprise at least one of:
 a temperature;
 a concentration; 
 a pressure; 
 a flow rate into the processing container; 
 a flow rate out of the processing container; and 
 a pH level. 
   
     
     
         4 . The method of  claim 1 , further comprising:
 providing data from one or more sensors responsive to one or more process parameters related to the cryopreservation or reanimation method.   
     
     
         5 . The method of  claim 4 , further comprising:
 controlling one or more characteristics of at least one of the two or more fluids flowing into and out of the processing container according to the data provided by the one or more sensors.   
     
     
         6 . The method of  claim 4 , wherein the one or more of conditions comprise at least one of:
 a temperature;
 a concentration; 
   a pressure;
 a flow rate into the processing container; 
 a flow rate out of the processing container; 
 a pH level; 
   a sphericity of the one or more oocytes, embryos or blastocysts; and   a volume of the one or more oocytes, embryos or blastocysts.   
     
     
         7 . The method of  claim 1 , wherein the flowing of the two or more fluids is controlled by a central controller adapted to cause flow through one or more valves. 
     
     
         8 . The method of  claim 1 , wherein the two or more fluids comprise:
 a first fluid including one or more cryoprotectant components; and   a second fluid including one or more dehydrating components.   
     
     
         9 . The method of  claim 8 , wherein a concentration of the cryoprotectant components of the two or more fluids flowing into the processing container is increased, and wherein a concentration of the dehydrating components of the two or more fluids flowing into the processing container is increased. 
     
     
         10 . The method of  claim 9 , wherein the concentration of the cryoprotectant components is increased from about 0.0 M to about 1.5 M, and wherein the concentration of the dehydrating components of the fluid flowing into the processing container is increased from about 0.0 M to about 0.3 M. 
     
     
         11 . The method of  claim 8 , wherein a concentration of the cryoprotectant components of the two or more fluids flowing into the processing container is decreased, and wherein a concentration of the dehydrating components of the two or more fluids flowing into the processing container is decreased. 
     
     
         12 . The method of  claim 11 , wherein the concentration of the cryoprotectant components is decreased from about 1.5 M to about 0.0 M, and wherein the concentration of the dehydrating components of the fluid flowing into the processing container is decreased from about 0.3 M to about 0.0 M. 
     
     
         13 . An apparatus for cryopreserving or reanimating of oocytes, embryos or blastocysts, comprising:
 a processing container configured to hold one or more oocytes, embryos or blastocysts therein, the processing container being further configured to allow fluid to flow into and out of the processing container;   one or more fluid reservoirs operatively connected to the processing container; and   a central controller adapted to control the flow of fluids from the one or more reservoirs into the processing container.   
     
     
         14 . The apparatus of  claim 13 , wherein the processing container includes a membrane, the membrane coupled to the processing container to retain the one or more oocytes, embryos or blastocysts within the processing container, and the membrane configured for passage of the fluid. 
     
     
         15 . The apparatus of  claim 14 , wherein the membrane includes a plurality of pores for passage of the fluid, the plurality of pores having a diameter no greater than about 100 μm. 
     
     
         16 . The apparatus of  claim 13 , wherein the processing container further comprises a removable basket, the removable basket configured to hold the one or more oocytes, embryos or blastocysts therein. 
     
     
         17 . The apparatus of  claim 13 , wherein the processing container comprises a canister, the canister including:
 at least one input port adapted for inflow of the fluid; and   at least output port adapted for outflow of the fluid.   
     
     
         18 . The apparatus of  claim 13 , wherein the processing container comprises a track, the track including:
 a hollow tube forming a loop, the hollow tube configured for conveying the one or more oocytes, embryos or blastocysts within;   a first port disposed on the hollow tube, the first port adapted for insertion into and extraction from the hollow tube the one or more oocytes, embryos or blastocysts;   a second port disposed on the hollow tube, the second port adapted for flowing the fluid into the hollow tube; and   at least one opening disposed on the hollow tube for discharge of the fluid from the hollow tube, the at least one opening configured to prohibit discharge of the one or more oocytes, embryos or blastocysts.   
     
     
         19 . The apparatus of  claim 13 , wherein the one or more fluid reservoirs includes a plurality of sub-compartments, each of the plurality of sub-compartments holding one or more fluids adapted for cryopreserving or reanimating of the one or more oocytes, embryos or blastocysts. 
     
     
         20 . A system for use in an oocyte or embryo cryopreserving or reanimating apparatus, comprising:
 an container configured to hold one or more oocytes, embryos or blastocysts therein, the container operatively connected to the cryopreserving or reanimating apparatus; and   one or more fluid reservoirs configured to operatively connect to the cryopreserving or reanimating apparatus, the one or more fluid reservoirs containing a volume of fluid adapted for cryopreservation or reanimation of the one or more oocytes, embryos or blastocysts.   
     
     
         21 . A fluid reservoir for use in an cryopreserving or reanimating apparatus, comprising:
 a volume configured to hold a quantity of one or more fluids adapted for cryopreserving or reanimating oocytes, embryos or blastocysts; and   a coupler configured to couple the fluid reservoir to one or more adapters disposed on the cryopreserving or reanimating apparatus.   
     
     
         22 . The fluid reservoir of  claim 21 , wherein the coupler includes one or more mating features adapted such that the fluid reservoir is couplable to only one of the one or more adapters. 
     
     
         23 . The fluid reservoir of  claim 22 , wherein the mating features are indicative of the one or more fluids within the volume of the fluid reservoir. 
     
     
         24 . The fluid reservoir of  claim 22 , further including electronic indicia readable by the cryopreserving or reanimating apparatus. 
     
     
         25 . The fluid reservoir of  claim 24 , wherein the electronic indicia is indicative of a fluid reservoir authorized for use with the cryopreserving or reanimating apparatus. 
     
     
         26 . A method of isolating a subset of oocytes, embryos or blastocysts for reanimation from a population of oocytes, embryos or blastocysts that have been cryopresreved with a cryopreserving apparatus comprising:
 storing data relating to one or more process parameters obtained from the cryopreserving apparatus during cryopreservation of one or more oocytes, embryos or blastocysts therein,
 associating the data with the one or more oocytes, embryos or blastocysts; 
 accessing the data; 
   analyzing the data to select a subset of oocytes, embryos or blastocysts for reanimation from a population of cryopreserved oocytes, embryos or blastocysts for which data has been stored.   
     
     
         27 . The method of  claim 26 , wherein the one or more process parameters comprise at least one of:
 a temperature;   a concentration;   a pressure;   a flow rate into the processing container;   a flow rate out of the processing container;   a pH level;   a sphericity of the one or more oocytes, embryos or blastocysts; and   a volume of the one or more oocytes, embryos or blastocysts.   
     
     
         28 . The method of  claim 26 , further comprising:
 generating a composite score based on at least two of the one or more cryopreservation parameters for the subset of oocytes, embryos or blastocysts,   wherein the composite score is indicative of the relative viability of the subset of oocytes, embryos or blastocysts.   
     
     
         29 . A central controller coupled to valves configured to control flow of fluids into and out of a processing container for oocytes, embryos or blastocysts and coupled to a thermal controller, the central controller being configured to:
 (a) introduce an aqueous solution into the processing container containing oocytes, embryos or blastocysts, the aqueous solution comprising a cryoprotectant maintained at a temperature of from about 33° to about 38° C. to replace at least a portion of intracytoplasmic water in the oocytes, embryos or blastocysts by the cryoprotectant;   (b) replace the aqueous solution in the processing container with a first dehydrating/cryoprotecting solution comprising a cryoprotectant and a dehydrating agent maintained at a temperature of from about 33° to about 38° C. to remove additional intra-cytoplasmic water while introducing additional cryoprotectant into the cytoplasm; and   (c) decrease the temperature of the dehydrating/cryoprotecting solution to a temperature of about 22° to about 26° C. during a period of from about 3 to 10 minutes.   
     
     
         30 . A computer program product, embodied on a computer-readable storage medium; for controlling valves configured to control flow of fluids into and out of a processing container for oocytes, embryos or blastocysts and for controlling a thermal controller, the computer program product comprising:
 (a) computer code for introducing an aqueous solution into the processing container containing oocytes, embryos or blastocysts, the aqueous solution comprising a cryoprotectant maintained at a temperature of from about 33° to about 38° C. to replace at least a portion of intracytoplasmic water in the oocytes, embryos or blastocysts by the cryoprotectant;   (b) computer code for replacing the aqueous solution in the processing container with a first dehydrating/cryoprotecting solution comprising a cryoprotectant and a dehydrating agent maintained at a temperature of from about 33° to about 38° C. to remove additional intra-cytoplasmic water while introducing additional cryoprotectant into the cytoplasm; and   (c) computer code for decreasing the temperature of the dehydrating/cryoprotecting solution to a temperature of about 22° to about 26° C. during a period of from about 3 to about 10 minutes.   
     
     
         31 . A central controller coupled to valves configured to control flow of fluids into and out of a processing container for oocytes, embryos or blastocysts and coupled to a thermal controller, the central controller being configured to:
 (a) introduce a first aqueous solution into the processing container containing cryopreserved oocytes, embryos or blastocysts, the aqueous solution comprising a cryoprotectant maintained at a temperature of from about 28° to about 35° C.; and   (b) replace the first aqueous solution in the processing container with a second aqueous solution comprising a cryoprotectant maintained at a temperature of from about 30° to about 34° C., wherein the concentration of cryoprotectant of the second aqueous solution is lower than the concentration of cryoprotectant of the first aqueous solution.   
     
     
         32 . The central controller of  claim 31 , further configured to:
 replace the second aqueous solution in the processing container with a third aqueous solution comprising a cryoprotectant maintained at a temperature of from about 30° to about 34° C., wherein the concentration of cryoprotectant of the third aqueous solution is lower than the concentration of cryoprotectant of the second aqueous solution.   
     
     
         33 . The central controller of  claim 31 , wherein the concentration of cryoprotectant of the third aqueous solution is less than about 0.1 M. 
     
     
         34 . A computer program product, embodied on a computer-readable storage medium; for controlling valves configured to control flow of fluids into and out of a processing container for oocytes, embryos or blastocysts and for controlling a thermal controller, the computer program product comprising:
 (a) computer code for introducing a first aqueous solution into the processing container containing cryopreserved oocytes, embryos or blastocysts, the aqueous solution comprising a cryoprotectant maintained at a temperature of from about 28° to about 35° C.; and   (b) computer code for replacing the first aqueous solution in the processing container with a second aqueous solution comprising a cryoprotectant maintained at a temperature of from about 30° to about 34° C., wherein the concentration of cryoprotectant of the second aqueous solution is lower than the concentration of cryoprotectant of the first aqueous solution.   
     
     
         35 . A method for evaluating a viability of a cryoprotected oocyte, the method comprising:
 (a) positioning a one or more oocytes in a cryoprotecting apparatus;   (b) monitoring a sphericity of the one or more oocytes during cryoprotection;   (c) measuring a degree of deviation from the sphericity of the one or more oocytes and evaluating a viability of a cryoprotected oocyte.   
     
     
         36 . The method of  claim 35 , wherein the cryoprotecting apparatus is of  claim 13 . 
     
     
         37 . The method of  claim 35 , wherein the monitoring is provided by a central controller. 
     
     
         38 . The method of  claim 35 , wherein the measurement is provided by the data stored by the central controller in the memory unit. 
     
     
         39 . A method of maturing an egg or developing an embryo, comprising:
 positioning an egg or an embryo in a processing container, the processing container configured to allow two or more fluids to flow into and out of the processing container;   flowing the two or more fluids into and out of the processing container with the egg or the embryo therein.   
     
     
         40 . The method of  claim 39 , further comprising:
 controlling one or more characteristics of at least one of the two or more fluids flowing into and out of the processing container according to one or more process parameters.   
     
     
         41 . The method of  claim 39 , wherein the flowing of the two or more fluids into and out of the processing container substantiality simulates an in vivo environment. 
     
     
         42 . The method of  claim 41 , wherein the egg or embryo experiences a simulated relative velocity from about 2 cm/day to about 1 cm/hour. 
     
     
         43 . An apparatus for maturing or developing an egg or an embryo, comprising:
 a processing container configured to hold an egg or an embryo therein, the processing container being further configured to allow fluid to flow into and out of the processing container;   one or more fluid reservoirs operatively connected to the processing container; and   a central controller adapted to control the flow of fluids from the one or more reservoirs into the processing container.   
     
     
         44 . The apparatus of  claim 43 , further comprising an imaging component capable of observing the one or more eggs and/or embryos in processing container. 
     
     
         45 . The apparatus of  claim 43 , wherein the processing container includes a membrane, the membrane coupled to the processing container to retain the egg or embryo within the processing container, and the membrane configured for passage of the fluid. 
     
     
         46 . The apparatus of  claim 43 , wherein the processing container comprises a canister, the canister including:
 at least one input port adapted for inflow of the fluid; and   at least output port adapted for outflow of the fluid.   
     
     
         47 . The apparatus of  claim 43 , wherein the processing container comprises a track, the track including:
 a hollow tube forming a loop, the hollow tube configured for conveying the egg or the embryo within;   a first port disposed on the hollow tube, the first port adapted for insertion into and extraction from the hollow tube the egg or embryo;   a second port disposed on the hollow tube, the second port adapted for flowing the fluid into the hollow tube; and   at least one opening disposed on the hollow tube for discharge of the fluid from the hollow tube, the at least one opening configured to prohibit discharge of the egg or the embryo.   
     
     
         48 . The apparatus of  claim 43 , wherein the one or more fluid reservoirs includes a plurality of sub-compartments, each of the plurality of sub-compartments holding one or more fluids adapted for maturing or developing the egg or the embryo. 
     
     
         49 . A system for use in an egg or an embryo maturing or developing apparatus, comprising:
 a container configured to hold one or more eggs and/or one or more embryos therein, the container operatively connected to the maturing or developing apparatus; and   one or more fluid reservoirs configured to operatively connect to the maturing or developing apparatus, the one or more fluid reservoirs containing a volume of fluid adapted for maturing or developing the one or more eggs or embryos.