System and sequential culture media for in vitro fertilization

Abstract

Instead of immersing human reproductive cells in a single culture medium throughout the various procedures used in IVF, a process is provided by which the reproductive cells may be moved through a sequence of distinct culture media as the various IVF procedures are carried out. In one implementation, the culture media specifically formulated to provide a physical environment similar to that found within the female reproductive tract and conducive to growth and development of human reproductive cells during the various stages of the IVF process. In this regard, specifically formulated culture media can be applied to support the reproductive cells in one or more of the following procedures: oocyte retrieval and handling; oocyte maturation; ordinary fertilization; oocyte, zygote and embryo examination and biopsy; embryonic development to the eight-cell stage; embryonic development to the blastocyst stage; embryo transfer, and cryopreservation.

1 . An in vitro fertilization medium supplement, comprising: alanyl-glutamine and one or more non-essential amino acids wherein the alanyl-glutamine and one or more non-essential amino acids are present in concentrations that support the development of a embryo cultured in a culture medium containing the medium supplement.  
     
     
         2 . The in vitro fertilization medium supplement of  claim 1 , wherein the one or more non-essential amino acids does not include taurine.  
     
     
         3 . An in vitro fertilization medium for supporting embryo development, comprising: 
 (a) water;    (b) a buffer present in an amount to buffer the in vitro fertilization medium within physiological pH;    (c) ionic constituents comprising sodium, potassium, calcium and magnesium; and    (d) alanyl-glutamine and one or more non-essential amino acids;    wherein the water, buffer, ionic constituents, alanyl-glutamine and one or more non-essential amino acids are present in concentrations that support the development of an embryo cultured in the medium    
     
     
         4 . The in vitro fertilization medium of  claim 3  further comprising one or more of glucose, lactate and pyruvate present in an amount sufficient to provide a source of energy for the embryo.  
     
     
         5 . The in vitro fertilization medium of  claim 3  wherein the ionic constituents further include phosphate.  
     
     
         6 . The in vitro fertilization medium of  claim 3  wherein the non-essential amino acids comprise one or more of alanine, aspartate, asparagine, glutamate, glycine, proline, serine and taurine.  
     
     
         7 . The in vitro fertilization medium of  claim 3  wherein the non-essential amino acids do not include taurine.  
     
     
         8 . The in vitro fertilization medium of  claim 3  further comprising one or more essential amino acids.  
     
     
         9 . The in vitro fertilization medium of  claim 3  further comprising one or more growth factors.  
     
     
         10 . The in vitro fertilization medium of  claim 9  wherein the growth factors are selected from insulin transferrin selenium, insulin-like growth factor and epidermal growth factor.  
     
     
         11 . The in vitro fertilization medium of  claim 3  further comprising an antibiotic.  
     
     
         12 . The in vitro fertilization medium of  claim 3  further comprising a hormone.  
     
     
         13 . The in vitro fertilization medium of  claim 3  further comprising one or more vitamins.  
     
     
         14 . The in vitro fertilization medium of  claim 3  further comprising EDTA.  
     
     
         15 . The in vitro fertilization medium of  claim 3  wherein EDTA is absent from the medium.  
     
     
         16 . The in vitro fertilization medium of  claim 3  wherein phosphate is absent from the medium.  
     
     
         17 . The in vitro fertilization medium of  claim 3  further comprising one or more of human serum albumin, hyaluronate, polyvinylpyrolidine, cysteamine, hyaluronidase and glutathione.  
     
     
         18 . The in vitro fertilization medium of  claim 3  wherein the buffer is selected from 4-morpholinepropanesulfonic acid, N-2-hydroxyethylpiperazine-N′-2 ethane sulfonic acid and bicarbonate.  
     
     
         19 . The in vitro fertilization medium of  claim 3  wherein bicarbonate is not used as a buffer.  
     
     
         20 . The in vitro fertilization medium of  claim 3  further comprising the embryo.  
     
     
         21 . The in vitro fertilization medium of  claim 3  wherein the medium supports embryonic development through the eight-cell stage.  
     
     
         22 . The in vitro fertilization medium of  claim 3  wherein the medium supports development of at least 84% of embryos cultured therein to the eight-cell stage.  
     
     
         23 . The in vitro fertilization medium of  claim 3  wherein the medium supports embryonic development from the eight-cell stage through the blastocyst stage.  
     
     
         24 . The in vitro fertilization medium of  claim 23  wherein the medium supports embryonic development of at least 77% of embryos cultured therein from the eight-cell stage to the blastocyst stage.  
     
     
         25 . The in vitro fertilization medium of  claim 3  wherein the medium further comprises at least one of lactate and pyruvate.  
     
     
         26 . The in vitro fertilization medium of  claim 3  wherein the medium further comprises glucose, lactate and pyruvate.  
     
     
         27 . The in vitro fertilization medium of  claim 3  further comprising an embryo that has been transferred from a medium that supports embryonic development following fertilization.  
     
     
         28 . The in vitro fertilization medium of  claim 3  wherein the embryo has been transferred at or after reaching the eight-cell stage.