Sperm cryoprotective media

Abstract

Described herein are methods for sperm cryoprotection and compositions comprising a cryoprotectant; a membrane protectant that stabilizes or assists in stabilization of membranes of sperm; and a free radical scavenger (e.g., a reducing agent, an antioxidant).

1 . A method for cryogenically preserving rodent sperm, comprising adding to the sperm, prior to cryopreservation, a composition that comprises at least one cryoprotectant; at least one membrane protectant; and reduced glutathione, beta-mercaptoethanol or a combination thereof. 
     
     
         2 . The method of  claim 1 , wherein the cryoprotectant is selected from the group consisting of raffinose, lactose, trehalose, melibiose, melezitose, mannotriose, stachyose, dextran, sucrose, and sugar alcohols thereof, glycerol, maltitol, and lactitol. 
     
     
         3 . The method of  claim 1 , wherein the cryoprotectant is raffinose or lactose. 
     
     
         4 . The method of  claim 1 , wherein the membrane protectant is a protein selected from the group consisting of egg protein, egg yolk protein, egg white protein, casein, albumin, keratin, collagen, atelocollagen, elastin, gelatin, peptones, fibrinogen, fibronectin, a soy protein, a wheat protein, a corn protein, a milk protein, and hydrolysates thereof. 
     
     
         5 . The method of  claim 1 , wherein the membrane protectant is skim milk or a component thereof milk powder or a component thereof; or egg yolk or a component thereof. 
     
     
         6 . The method of  claim 1 , wherein the rodent sperm is isolated from a mouse. 
     
     
         7 . The method of  claim 6 , wherein the mouse is an inbred mouse strain or substrain. 
     
     
         8 . The method of  claim 1 , wherein the rodent sperm is isolated from a rat. 
     
     
         9 . The method of  claim 1 , wherein the cryoprotectant is raffinose, lactose, or glycerol; the membrane protectant is milk, milk protein, a protein in milk, egg protein, egg yolk or egg white protein. 
     
     
         10 . The method of  claim 1 , wherein the at least one cryoprotectant comprises at least one sugar. 
     
     
         11 . The method of  claim 10 , wherein the sugar is raffinose or lactose; and the membrane protectant is egg yolk, skim milk, milk or a component thereof. 
     
     
         12 . The method of  claim 1 , wherein the composition further comprises at least one of: monothioglycerol, dithiothreitol, Tris(2-carboxyethyl) phosphine, dithioerythritol, thioredoxin, dithionite, 2-mercaptoethylamine, dimethyl thiourea, DMSO, nordihydroguaiaretic acid (NDGA), 2,3-dimercapto-1-propanol, hydroquinone, an amino acid, an amino acid derivative, a vitamin, a vitamin derivative, butylated hydroxyanisole, butylated hydroxytoluene, dihydrolipoic acid, tetrahydropapaveroline, 2-thiobarbituric acid or taurine, dimercaptosuccinic acid, allopurinol, deferoxamine, melatonin, catalase, glutathione peroxidase, superoxide dismutase, steroids; or a combination of any two or more thereof. 
     
     
         13 . A method for cryogenically preserving rodent sperm, comprising adding to the rodent sperm, prior to cryopreservation, a composition that includes at least one cryoprotectant; and reduced glutathione, beta-mercaptoethanol or a combination thereof. 
     
     
         14 . The method of  claim 13 , wherein the cryoprotectant is selected from the group consisting of raffinose, lactose, trehalose, melibiose, melezitose, mannotriose, stachyose, dextran, sucrose, and sugar alcohols thereof, glycerol, maltitol, and lactitol. 
     
     
         15 . The method of  claim 13 , wherein the composition further comprises at least on of: monothioglycerol, dithiothreitol, Tris(2-carboxyethyl) phosphine, dithioerythritol, thioredoxin, dithionite, 2-mercaptoethylamine, dimethyl thiourea, DMSO, nordihydroguaiaretic acid (NDGA), 2,3-dimercapto-1-propanol, hydroquinone, an amino acid, an amino acid derivative, a vitamin, a vitamin derivative, butylated hydroxyanisole, butylated hydroxytoluene, dihydrolipoic acid, tetrahydropapaveroline, 2-thiobarbituric acid or taurine, dimercaptosuccinic acid, allopurinol, deferoxamine, melatonin, catalase, glutathione peroxidase, superoxide dismutase, steroids; or a combination of any two or more thereof. 
     
     
         16 . The method of  claim 13 , wherein the rodent sperm is isolated from a mouse. 
     
     
         17 . The method of  claim 16 , wherein the mouse is an inbred mouse strain or substrain. 
     
     
         18 . The method of  claim 13 , wherein the rodent sperm is isolated from a rat. 
     
     
         19 . A method for cryogenically preserving rodent sperm, comprising adding to the rodent sperm, prior to cryopreservation, a composition that includes reduced glutathione, beta-mercaptoethanol or a combination thereof. 
     
     
         20 . The method of  claim 19 , further comprising at least one of: monothioglycerol, dithiothreitol, Tris(2-carboxyethyl) phosphine, dithioerythritol, thioredoxin, dithionite, 2-mercaptoethylamine, dimethyl thiourea, DMSO, nordihydroguaiaretic acid (NDGA), 2,3-dimercapto-1-propanol, hydroquinone, an amino acid, an amino acid derivative, a vitamin, a vitamin derivative, butylated hydroxyanisole, butylated hydroxytoluene, dihydrolipoic acid, tetrahydropapaveroline, 2-thiobarbituric acid or taurine, dimercaptosuccinic acid, allopurinol, deferoxamine, melatonin, catalase, glutathione peroxidase, superoxide dismutase, a steroid; or a combination of any two or more thereof.