Abnormal syngamy phenotypes observed with time lapse imaging for early identification of embryos with lower development potential

• US 10,241,108 B2
• Filed: 02/03/2014
• Issued: 03/26/2019
• Est. Priority Date: 02/01/2013
• Status: Active Grant

• Current Assignee: Ares Trading SA (Merck & Co., Inc.)
• Original Assignee: Ares Trading SA (Merck & Co., Inc.)
• Inventors: Shen, Shehua, Chen Kim, Alice A., Wirka, Kelly Athayde, Suraj, Vaishali, Tan, Lei
• Primary Examiner(s): Moloye, Titilayo
• Application Number:
  US14/171,610
• Publication Number:
  US 20140220619A1
• Time in Patent Office: 1,877 Days
• CPC Class Codes: G01N 2800/367 Infertility, e.g. sperm disorder, ovulatory dysfunction
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism

Abstract

Methods, compositions and kits for determining the developmental potential of one or more embryos are provided. These methods, compositions and kits find use in identifying embryos in vitro that are most useful in treating infertility in humans.

Claims

1. A method for treating infertility in a female human subject, comprising:

(A) providing one or more human embryos under in vitro culture conditions sufficient for embryo development;

(B) imaging said one or more human embryos for a time period from at least syngamy to at least a first cytokinesis to create images, wherein said one or more human embryos comprise at least one cellular parameter;

(C) identifying one or more abnormal syngamy (AS) embryos when said time period is less than 2.5 hours and one or more normal syngamy (NS) embryos when said time period is greater than 2.5 hours;

(D) deselecting said one or more human embryos when at least one of said images comprises said one or more AS embryos;

(E) selecting said one or more human embryos when at least one of said images comprises said one or more NS embryos; and

(F) developing at least one of said selected NS embryos under said in vitro culture conditions into a normal human blastocyst embryo; and

(G) treating infertility of a female human subject with said normal human blastocyst embryo by in vitro fertilization.

2. The method of claim 1 wherein said at least one cellular parameter is selected from the group consisting of:

(a) the duration of the first cytokinesis;

(b) the time between the first and second mitosis;

(c) the time between the second and third mitosis;

(d) the time interval between cytokinesis 1 and cytokinesis 2;

(e) the time interval between cytokinesis 2 and cytokinesis 3;

(f) the time interval between fertilization and the 5 cell stage; and

(g) the duration of the first cell cycle.

3. The method of claim 1 wherein said one or more human embryos are produced by fertilization of oocytes in vitro.

4. The method of claim 3 wherein said oocytes are matured in vitro.

5. The method of claim 4 wherein said oocytes matured in vitro are supplemented with growth factors.

6. The method of claim 1 wherein said one or more human embryos have not been frozen prior to said providing.

7. The method of claim 1 wherein said one or more human embryos have been frozen prior to said providing.

8. The method of claim 1 wherein said selecting of said normal human embryo is automated.

9. The method of claim 1 wherein said images are digitally stored.

10. The method of claim 1 wherein said imaging employs darkfield illumination.

11. The method of claim 1 wherein said one or more human embryos are provided in a culture dish.

12. The method of claim 11 wherein said culture dish comprises a plurality of microwells.

13. The method of claim 12 wherein said culture dish comprises up to about 30 microwells.

14. The method of claim 12 wherein said one or more human embryos is provided in at least one of said plurality of microwells.

15. The method of claim 1 wherein the imaging is carried out at an imaging station.

16. The method of claim 1, wherein said AS embryo further comprises said at least one cellular parameter selected from the group consisting of a disordered pronuclei movement, a delayed dispersion of nuclear envelopes, and an active oolema movement before the dispersion of the nuclear envelopes.

17. The method of claim 1, wherein said NS embryo further comprises said at least one cellular parameter selected from the group consisting of a timely disappearance of pronuclei, a smooth dispersion of the nuclear envelopes, a minimal or no pronuclear movement within the cytoplasm, a minimal or no oolema movement before the dispersion of nuclear envelopes and a first cytokinesis duration is about 0-33 minutes.