Culture media for developmental cells containing elevated concentrations of lipoic acid

Abstract

A composition and method for in vitro fertilization is provided which uses culture media comprising elevated concentrations of lipoic acid. More specifically, the invention provides culture media for developmental cells having a lipoic acid concentration of 5 μM to 40 μM. Culture media that include lipoic acid at concentrations within the identified range are able to provide blastocysts with increased survival, increased cell numbers, increased inner cell masses and/or increased percentage of the total mass made up by the inner cell compared to blastocysts cultured in a control medium.

1 . A developmental cell culture medium comprising:
 (a) water;   (b) inorganic salts;   (c) at least one energy source; and   (d) lipoic acid, lipoic acid derivatives or a combination thereof at a concentration of about 5 μM to about 40 μM.   
     
     
         2 . The culture medium of  claim 1  further comprising one or more amino acids. 
     
     
         3 . The culture medium of  claim 1 , wherein the culture medium supports the development of at least one of stem cells, gametes, zygotes or embryos from one developmental stage to another developmental stage. 
     
     
         4 - 8 . (canceled) 
     
     
         9 . The culture medium of  claim 1 , further comprising one or more components selected from the group consisting of growth factors, hormones, antibiotics, and vitamins. 
     
     
         10 . The culture medium of  claim 1 , further comprising ethanol. 
     
     
         11 . The culture medium of  claim 1 , having a lipoic acid concentration of about 5 μM to about 20 μM. 
     
     
         12 . The culture medium of  claim 1 , having a lipoic acid concentration of about 5 μM to about 10 μM. 
     
     
         13 . The culture medium of  claim 1 , having a lipoic acid concentration of about 8 μM to about 12 μM. 
     
     
         14 . The culture medium of  claim 1 , having a lipoic acid concentration of about 10 μM. 
     
     
         15 - 19 . (canceled) 
     
     
         20 . A method for culturing developmental cells, the method comprising culturing a stem cell, gamete, zygote or embryo in the culture medium of  claim 1 . 
     
     
         21 . The method of  claim 20 , wherein the stem cell, gamete, zygote or embryo is cultured under ambient oxygen conditions. 
     
     
         22 . The method of  claim 20 , wherein the stem cell, gamete, zygote or embryo is cultured under reduced oxygen conditions. 
     
     
         23 . The method of  claim 20 , wherein the step of culturing a stem cell, gamete, zygote, or embryo comprises culturing a human embryo. 
     
     
         24 . The method of  claim 20 , wherein the step of culturing a stem cell, gamete, zygote or embryo comprises culturing an embryo up to the 4-cell stage. 
     
     
         25 . The method of  claim 20 , wherein the step of culturing a stem cell, gamete, zygote or embryo comprises culturing an embryo up to the 8-cell stage. 
     
     
         26 . The method of  claim 20  wherein the step of culturing a stem cell, gamete, zygote or embryo comprises culturing an embryo up to the blastocyst stage. 
     
     
         27 . The method of  claim 20 , wherein the step of culturing a stem cell, gamete, zygote or embryo comprises culturing an embryo from the 8-cell stage to the blastocyst stage. 
     
     
         28 . The method of  claim 20 , wherein the step of culturing a stem cell, gamete, zygote or embryo comprises culturing an embryo to the 3 day stage. 
     
     
         29 . The method of  claim 20 , wherein the step of culturing a stem cell, gamete, zygote or embryo comprises culturing an embryo to the 5 day stage. 
     
     
         30 . A developmental cell culture medium comprising:
 (a) water;   (b) inorganic salts;   (c) at least one energy source; and   (d) lipoic acid, lipoic acid derivatives or a combination thereof, wherein the culture medium supports the development of mammalian embryos to the blastocyst stage, and further wherein the culture medium provides at least a 10% increase in the total number of blastocysts at day 5 compared to a control medium that differs from the culture medium only in the absence of lipoic acid and lipoic acid derivatives.   
     
     
         31 . The medium of  claim 30  further comprising one or more amino acids. 
     
     
         32 . The medium of  claim 30 , wherein the mammalian embryos are CF1 mouse embryos. 
     
     
         33 . The medium of  claim 30 , wherein the developmental cell culture medium comprises 5 μM to 40 μM of lipoic acid, lipoic acid derivatives or a combination thereof. 
     
     
         34 . A method for culturing embryos, the method comprising culturing an embryo in the culture medium of  claim 30  to the blastocyst stage. 
     
     
         35 . A developmental cell culture medium comprising:
 (a) water;   (b) inorganic salts;   (c) at least one energy source; and   (d) lipoic acid, lipoic acid derivatives or a combination thereof;   
       wherein the culture medium supports the development of mammalian embryos to the blastocyst stage, and further wherein the culture medium provides at least a 10% increase in the total cell number of the blastocysts at day 5 compared to a control medium that differs from the culture medium only in the absence of lipoic acid and lipoic acid derivatives. 
     
     
         36 . The medium of  claim 33  further comprising one or more amino acids. 
     
     
         37 . The medium of  claim 33 , wherein the mammalian embryos are CF1 mouse embryos. 
     
     
         38 . The medium of  claim 33 , wherein the developmental cell culture medium comprises 5 μM to 40 μM of lipoic acid, lipoic acid derivatives or a combination thereof 
     
     
         39 . A method for culturing embryos, the method comprising culturing an embryo in the culture medium of  claim 33  to the blastocyst stage. 
     
     
         40 . A developmental cell culture medium comprising:
 (a) water;   (b) inorganic salts;   (c) at least one energy source; and   (e) lipoic acid, lipoic acid derivatives, or a combination thereof;   
       wherein the culture medium supports the development of mammalian embryos to the blastocyst stage, and further wherein the culture medium provides at least a 10% increase in the inner cell mass of the blastocysts at day 5 compared to a control medium that differs from the culture medium only in the absence of lipoic acid and lipoic acid derivatives. 
     
     
         41 . The medium of  claim 37  further comprising one or more amino acids. 
     
     
         42 . The medium of  claim 37 , wherein the mammalian embryos are CF1 mouse embryos. 
     
     
         43 . The medium of  claim 37 , wherein the developmental cell culture medium comprises 5 μM to 40 μM of lipoic acid, lipoic acid derivatives or a combination thereof 
     
     
         44 . A method for culturing embryos, the method comprising culturing an embryo in the culture medium of  claim 37  to the blastocyst stage. 
     
     
         45 . The culture medium of  claim 2 , wherein the concentration of inorganic salts in the medium is about 115 mM to about 140 mM, the concentration of energy sources in the medium is about 5 mM to about 30 mM, and the total concentration of amino acids in the medium is about 0.1 to 15 mM. 
     
     
         46 . The culture medium of  claim 1 , further comprising a chelating agent, an antibiotic and hyaluronan.