Imaging and evaluating embryos, oocytes, and stem cells
Methods, compositions and kits for determining the developmental potential of one or more embryos or pluripotent cells and/or the presence of chromosomal abnormalities in one or more embryos or pluripotent cells are provided. These methods, compositions and kits find use in identifying embryos and oocytes in vitro that are most useful in treating infertility in humans.
1. A method for selecting a human embryo comprising:
(a) culturing a human embryo in vitro under conditions sufficient for embryonic development;
(b) imaging said human embryo by time lapse microscopy wherein images are captured at time intervals necessary to detect one or more cellular parameters comprising, the time interval between cytokinesis 1 and cytokinesis 2;
(c) analyzing said images to arrive at a measurement of said one or more cellular parameters;
(d) assessing the developmental potential of the human embryo from said one or more cellular parameter measurements to determine if the embryo has good developmental potential or poor developmental potential, wherein the human embryo is determined to have good developmental potential when the time interval between cytokinesis 1 and cytokinesis 2 is about 8-15 hours or poor developmental potential when the time interval between cytokinesis 1 and cytokinesis 2 lasts more than about 15 hours or less than about 8 hours; and
(e) selecting an embryo with a good developmental potential for transfer into a female recipient or selecting an embryo with poor developmental potential for nontransfer.
2. The method of claim 1 wherein said one or more cellular parameters further comprises one or more of cell size, thickness of the zona pellucida, degree of fragmentation, and symmetry of daughter cells resulting from a cell division.
3. The method of claim 1 wherein one or more cellular parameter further comprises one or more of the duration of the second cytokinesis, the duration of the 3rd cytokinesis, the duration of the 4th cytokinesis, the duration of cell cycle 1, the duration of cell cycle 2, the duration of cell cycle 3, and the duration of cell cycle 4.
4. The method of claim 1 wherein the one or more cellular parameters further comprises one or more of the interval between initiation of cytokinesis 1 and the initiation of cytokinesis 2, the interval between the resolution of cytokinesis 1 and the resolution of cytokinesis 2, the interval between the initiation of cytokinesis 1 and the resolution of cytokinesis 2; the interval between the resolution of cytokinesis 1 and the initiation of cytokinesis 2; the initiation of cytokinesis 2 and the initiation of cytokinesis 3, the interval between resolution of the cytokinesis 2 and the resolution of cytokinesis 3, the interval between initiation of cytokinesis 2 and the resolution of cytokinesis 3, the interval between resolution of cytokinesis 2 and the initiation of cytokinesis 3.
5. The method of claim 1 wherein the cellular parameters are measured manually.
6. The method of claim 1 wherein the cellular parameters are measured automatically.
7. The method of claim 1 wherein images are captured every 5 minutes.
8. The method of claim 7 wherein images are captured at 0.5 seconds of light exposure per image.
9. The method of claim 1 wherein said measurements take place within 54 hours of fertilization.
10. The method of claim 1 wherein the embryo being assessed is a 5 cell embryo.
11. The method of claim 1 wherein said embryo is produced by fertilization of an oocyte in vitro.
12. The method of claim 11 wherein said oocyte is matured in vitro.
13. The method of claim 12 wherein said oocyte matured in vitro is supplemented with growth factors.
14. The method of claim 1 wherein said embryo has not been frozen prior to the culturing under conditions sufficient for embryonic development.
15. The method of claim 1 wherein said embryo has been frozen prior to culturing.
16. The method of claim 1 further comprising transferring a human embryo determined to have good developmental potential into a female recipient.
17. The method of claim 1 wherein the images are digitally stored.
18. The method of claim 1 wherein said time lapse microscopy employs darkfield illumination.
19. The method of claim 1 wherein said human embryo is placed in a culture dish prior to culturing.
20. The method of claim 19 wherein said culture dish comprises a plurality of microwells.
21. The method of claim 20 wherein said culture dish comprises from 1 to about 30 microwells.
22. The method of claim 20 wherein the human embryo is placed within a microwell prior to culturing.