Vitro maturation of immature human oocytes

a technology of immature human oocytes and vitro maturation, which is applied in the field of assisted reproductive technology, can solve the problems of ovulation hyperstimulation syndrome (ohss), not all ovulations have matured to the m-ii stage, and the procedure of ovarian stimulation requires frequent blood sampling and ultrasound monitoring, and/or death

Inactive Publication Date: 2005-09-22
MCGILL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cumulus cells interfere with ICSI and obscure microscopic observation of the oocyte.
Unfortunately, not all of the oocytes have matured to the M-II stage when they are harvested in ovarian stimulation.
These procedures of ovarian stimulation require frequent blood sampling and ultrasound monitoring, at considerable cost.
Moreover, current IVF treatment causes substantial discomfort and, in some cases, results in ovarian hyperstimulation syndrome (OHSS) and / or even death.
There is also anxiety that the long-term effects of repeated ovarian stimulation may increase the risk of ovarian, endometrial and breast cancer.
Although the first pregnancy and live birth from immature oocytes retrieved from women with PCOS followed by IVM has been reported, the success rates are still low because oocyte maturation rates are relatively low.
However, oocyte maturation in vitro is profoundly affected by culture conditions.

Method used

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  • Vitro maturation of immature human oocytes
  • Vitro maturation of immature human oocytes
  • Vitro maturation of immature human oocytes

Examples

Experimental program
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Effect test

example 1

Source of Immature Oocytes

[0108] A total of 521 immature oocytes (213 oocytes at GV stage and 308 oocytes at M-I stage) were collected from 183 women (age range between 28 to 40 years) who underwent controlled ovarian stimulation and ICSI cycles under informed consent. Ovarian stimulation was performed with GnRHa, HMG or FSH and HCG using standard protocols.

[0109] 36 hours after HCG injection, oocyte retrieval was carried out using an ultrasound-guided trans-vaginal probe. The collected COC were cultured in human tubal fluid medium (HTF; Irvine Scientific) supplemented with 10% synthetic serum substitute (SSS; Irvine Scientific) and placed in a 5% CO2 incubator (37° C. with high humidity) for at least 1 h before they were stripped from their cumulus cells.

[0110] COC were stripped with 85 IU / ml hyaluronidase in HTF medium and mechanical pipetting. All oocytes were completely denuded from their cumulus cells. Denuded oocytes were observed under an inverted microscope to assess for...

example 2

[0135] A total of 40 women with PCOS underwent 40 completed IVM treatment cycles. All patients were under 40 years of age (mean 32.3±3.9) and some patients presented irregular menstrual cycles or anovulation. All patients had a minimum 2-year history of infertility.

[0136] If the patients had irregular menstrual cycles, the treatment cycles was initiated by the administration of intra-vaginal progesterone (Prometrium; Schering, Pointe-Clair, Quebec, Canada) in a dose of 300 mg daily for 10 days. Withdrawal bleeding occurred within 3 days after the last dose.

[0137] On day 2 or 3 following the onset of menstrual bleeding, the patients underwent a baseline ultrasound scan to ensure that no ovarian cysts were present. Transvaginal ultrasound scans were repeated on day 8 to exclude the development of a dominant follicle. The size of all follicles on ultrasound scan had to be <10 mm in diameter on day 8 of the cycle.

[0138] Oocyte retrieval was performed on day 10 to 14 of the cycle. Bef...

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Abstract

The invention provides methods for in vitro maturation of immature human oocytes. In one aspect, the invention provides a method for in vitro maturation of immature human oocytes, comprising the steps of: (a) inducing in a female human subject an increase in endogenous luteinizing hormone levels, said subject having not undergone an ovarian stimulation protocol prior to said inducing step; (b) obtaining from said subject an immature oocyte; and (c) culturing said oocyte until maturity.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of U.S. provisional patent application No. 60 / 362,395, filed Mar. 8, 2002, which is incorporated herein by reference.FIELD OF THE INVENTION [0002] The present invention relates to the field of assisted reproductive technology (ART), particularly to methods for culturing immature human oocytes so that they mature to metaphase-II stage capable of being fertilized and forming a viable embryo. BACKGROUND OF THE INVENTION [0003] Since the first successful human pregnancy by way of in vitro fertilization (IVF) was achieved in 1978, ART has helped thousands of thousands of women to overcome infertility problem [0004] Normally, during the follicular phase of a woman's reproductive cycle, only a single follicle grows to the preovulatory stage and releases its oocyte for potential fertilization. However, current IVF treatment requires that multiple oocytes be harvested. Therefore, women are normally pre-treated...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/25C12N2500/32C12N2500/38C12N2517/10C12N2501/115C12N2501/31C12N2501/39C12N2501/392C12N2501/11
Inventor CHIAN, RI-CHENG
Owner MCGILL UNIV
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