Method and quality control molecular based mouse embryo assay for use with in vitro fertilization technology

1. A method for assessing a product or culture condition for use in human Assisted Reproductive Technologies (ART), the method comprising:

(i) microscopically evaluating the fluorescence intensity or light emission of at least one fluorescent reporter gene operably linked to a regulatory region of OCT-4 during at least one stage of development of a transgenic murine test embryo,

wherein the test embryo comprises at least one cell and has been cultured in vitro in contact with the product or culture condition for a specified duration, and wherein the stage is a developmental stage between 1-cell zygote through blastocyst; and

(ii) determining the acceptability of the product or culture condition for use in human ART based upon the evaluation, wherein the product or culture condition is deemed acceptable when the fluorescence intensity or light emission of the fluorescent reporter gene in the test embryo is greater than a threshold level of the fluorescence intensity or light emission and/or localization of fluorescence of a reporter gene in a control embryo.

2. The method of claim 1 , further comprising evaluating the morphology of the embryo during at least one stage of development of said embryo.

3. The method of claim 1 , wherein the transgenic embryo is a one-cell embryo or a two-cell embryo.

4. The method of claim 1 , wherein the at least one fluorescent reporter gene encodes a fluorescent protein selected from the group consisting of green fluorescent protein, red fluorescent protein, cyan fluorescent protein, orange fluorescent protein, yellow fluorescent protein.

5. The method of claim 1 , wherein said evaluating step comprises:

(i) capturing at least one image of said embryo;

(ii) determining at least one of fluorescence intensity or light emission and/or localization of fluorescence of said reporter gene based on the image.

6. The method of claim 5 , wherein step (ii) is performed by a computer.

7. The method of claim 1 , wherein the control embryo has been cultured in the absence of the product or culture condition.

8. The method of claim 1 , wherein the at least one stage of development is 1-cell stage, 2-cell stage, 4-cell stage, 8-cell stage, morula stage, and/or blastocyst stage.

9. The method of claim 1 , wherein the product is selected from the group consisting of needles, catheters, microtools, labware, syringes, tissue culture dishes, tissue culture plates, pipette tips, dishes, plates, water, water purification systems, gamete and embryo culture media, gamete and embryo handling/processing media, transport media, enzymes for denuding oocytes, gradient for sperm separation, freezing/vitrification media, thawing/warming media, pipette and embryo handling devices, lab-ware used in the process of human ART including but not limited to Petri dishes, centrifuge tubes, cryopreservation and cryo-storage devices, and any solutions, reagents or devices involved with ART, media supplements, and other vessels, devices, or reagents that come into physical contact with gametes, embryos or tissue culture media.

10. The method of claim 1 , wherein the control embryo has been cultured with a control product.

11. The method of claim 1 , wherein the fluorescence intensity or light emission of the fluorescent reporter gene is greater than a threshold level in a given location of the cell to indicate that the test embryo is not affected by use of the product or culture condition when the reporter gene is expressed at or above a threshold level and in the location.

12. The method of claim 11 , wherein the fluorescent reporter gene is normally expressed in the cytoplasm at a time when fluorescence intensity or light emission is evaluated, and the conditions are deemed optimal when the fluorescence intensity or light emission in the cytoplasm of the test embryo is higher than the threshold level of the fluorescence intensity or light emission in the cytoplasm of the control embryo.

13. The method of claim 11 , wherein the fluorescent reporter gene is normally expressed in the nucleus at a time when fluorescence is evaluated, and the conditions are deemed optimal when the fluorescence in the nucleus of the test embryo is higher than the threshold level of fluorescence in the nucleus of the control embryo.

14. The method of claim 1 , wherein the threshold level comprises at least one of 50% or greater, 60% or greater, 70% or greater, 80% or greater, 90% or greater, or 100% or greater of the fluorescence intensity or the light emission of a reporter gene observed in a given location of the control embryo.

15. The method of claim 14 , wherein a plurality of test embryos are cultured and the conditions are deemed suboptimal when the fluorescence from less than 80% of the test embryos is equal to or greater than the threshold level of at least one of 50% or greater, 60% or greater, 70% or greater, 80% or greater, 90% or greater, or 100% or greater fluorescence intensity or light emission from the control embryo.

16. The method of claim 15 , wherein suboptimal culture conditions indicate that the product is not suitable for use in human ART.

17. The method of claim 1 , wherein fluorescence intensity is ranked on a scale wherein a score of 0 indicates no fluorescence intensity, a score of 1 indicates low fluorescence intensity, a score of 2 indicates medium fluorescence intensity, and a score of 3 indicates high fluorescence intensity.

18. The method of claim 17 , comprising culturing a plurality of test embryos under the specified conditions, wherein the conditions are deemed suboptimal when less than 60% of the test embryos exhibit fluorescence at a medium to high level.

19. The method of claim 17 , wherein acceptability of the product or culture condition is dependent on from about 60% to about 100% of the test embryos exhibiting fluorescence intensity at or above the fluorescence intensity of the control embryos, wherein the control embryos have a score of 2 or above.