Methods and devices for generation of oocytes with improved oocyte quality for in vitro fertilization procedures using non-invasive cellular transfer

A technology of oocytes and mother cells, applied in the direction of epidermal cells/skin cells, non-embryonic pluripotent stem cells, biochemical equipment and methods, etc., to achieve the effect of improving the quality of oocytes

Pending Publication Date: 2021-08-03
THE CHINESE UNIVERSITY OF HONG KONG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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However, the existence of the

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  • Methods and devices for generation of oocytes with improved oocyte quality for in vitro fertilization procedures using non-invasive cellular transfer
  • Methods and devices for generation of oocytes with improved oocyte quality for in vitro fertilization procedures using non-invasive cellular transfer
  • Methods and devices for generation of oocytes with improved oocyte quality for in vitro fertilization procedures using non-invasive cellular transfer

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0126] Effect of Tunneling Nanotube-Forming Cells on Mouse Oocytes Using a Hanging Drop Culture System

[0127] The effect of transferring autologous biomolecules and cellular components to mouse oocytes using the hanging drop method during in vitro maturation was determined. C57 / BL6 mice aged 6 weeks or 12 months were perfused with PMSG (5 IU / female). Female mice were sacrificed 48 hours later by cervical dislocation. Squirt alcohol into the abdomen of the mouse, and make an incision to expose the abdominal cavity. The ovary was dissected with a needle and cut into small pieces. Blastocyst (GV) oocytes were selected and either controlled (medium only) or co-cultured with adult stem cells. Before co-cultivation, cumulus cells around oocytes were removed using 1 mg / μl hyaluronidase in M2 medium (Sigma-Aldrich, USA).

[0128] To mimic poor oocyte quality (such as by inhibiting mitochondrial respiration), oocytes were treated with rotenone at a concentration of 0.05 μΜ for ...

example 2

[0140] Effects of KIF5B overexpression in adult stem cells on mouse oocytes using a hanging drop culture system ring

[0141] The effect of autologous transfer of biomolecules and cellular components from adult stem cells overexpressing the KIF5B gene to mouse oocytes was determined using the hanging drop method. The KIF5B gene was cloned into an expression vector plasmid, and the plasmid was transfected into human adult stem cells.

[0142] C57 / BL6 mice were perfused with PMSG (5 IU / female). Female mice were sacrificed 48 hours later by cervical dislocation. Squirt alcohol into the abdomen of the mouse, and make an incision to expose the abdominal cavity. The ovary was dissected with a needle and cut into small pieces. Blastocyst (GV) oocytes were selected and either controlled (medium only) or co-cultured with adult stem cells. Before co-cultivation, cumulus cells around oocytes were removed using 1 mg / μl hyaluronidase in M2 medium (Sigma-Aldrich, USA). To simulate ...

example 3

[0149] Effect of adult stem cells on mouse oocytes using the microfluidic device of the present invention

[0150] The effect of using the microfluidic device of the present invention to transfer autologous biomolecules and cellular components from adult stem cells to mouse oocytes was determined in vitro. Six-week-old C57 / BL6 mice were perfused with PMSG (5 IU / female). Female mice were sacrificed 48 hours later by cervical dislocation. Squirt alcohol into the abdomen of the mouse, and make an incision to expose the abdominal cavity. The ovary was dissected with a needle and cut into small pieces. Blastocyst (GV) oocytes were selected and either controlled (medium only) or co-cultured with adult stem cells. Cumulus cells around oocytes were removed using 85 IU / ml hyaluronidase in M2 medium (Sigma-Aldrich, USA) before co-cultivation. To mimic poor oocyte quality (such as by inhibiting mitochondrial respiration), oocytes were treated with rotenone at a concentration of 0.0...

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Abstract

The invention pertains to methods and devices for generating mature oocytes from immature oocytes and improving oocyte quality for improved success of assistant reproductive technology (ART). The methods include the use of tunneling nanotube-forming cells and oocytes, wherein the tunneling nanotube-forming cells transfer autologous genomic materials, biomolecules and cellular components to the oocytes. The devices of the invention include micro fluidic device to improve the efficiency of the transfer of biomolecules and cellular components between tunneling nanotube-forming cells and oocytes.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Patent Application Serial No. 62 / 967,672, filed January 30, 2020, the entire contents of which, including any tables, figures, or drawings, are incorporated herein by reference. [0003] Background of the invention [0004] In vitro fertilization (IVF) is a technique used in assisted reproductive technology to increase fertility. Mature oocytes and sperm are collected from a donor, and the oocytes are fertilized in a laboratory. The fertilized egg is then implanted back into the donor's uterus. Collection of human oocytes requires controlled ovarian hyperstimulation and transvaginal oocyte retrieval. Controlled ovarian hyperstimulation primarily involves injections of gonadotropins such as follicle stimulating hormone followed by human chorionic gonadotropin (hCG) 36 hours prior to oocyte retrieval. Depending on the regimen, injections are usually supplemented with a gonadotr...

Claims

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Application Information

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IPC IPC(8): C12N5/075C12N5/0789C12N5/0775C12N5/0797C12N5/074C12N5/078B01L3/00
CPCB01L3/5027B01L2300/0861C12N5/0609C12N2502/088C12N2502/09C12N2502/11C12N2502/1171C12N2502/1352C12N2502/1382C12N2501/31C12N2502/03C12N2502/137C12M23/16C12M29/26
Inventor 李天立李天照陈耀樑李颖彤吴健颍陈霆羲
Owner THE CHINESE UNIVERSITY OF HONG KONG
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