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

A method for generating mature oocytes from immature oocytes, the method comprising:

• culturing one or more immature oocyte in IVM culture medium,
• culturing one or more tunneling nanotube-forming cell under standard culture conditions;
• transfecting the one or more tunneling nanotube-forming cell with a vector carrying a microtubule motor protein kinesin family member 5B (KIF5B) gene;
• combining the one or more immature oocyte and the one or more transfected tunneling nanotube-forming cell in a drop of culture medium, and
• suspending said drop of culture medium from an inverted surface, such that said one or more immature oocyte and said one or more tunneling nanotube-forming cell are in close proximity in the hanging drop and at least one tunneling nanotube-forming cell forms at least one tunneling nanotube to at least one immature oocyte and transfers biomolecules, cellular components, and/or organelles to the at least one immature oocyte through the at least one tunneling nanotube protruding from said at least one tunneling nanotube-forming cell for a period of at least one hour.

A method for generating mature oocytes from immature oocytes, the method comprising:

• culturing one or more immature oocyte in IVM culture medium,
• culturing one or more tunneling nanotube-forming cell under standard culture conditions;
• transfecting the one or more tunneling nanotube-forming cell with a vector carrying a microtubule motor protein kinesin family member 5B (KIF5B) gene;
• injecting said one or more oocyte and said one or more transfected tunneling nanotube-forming cell into a microfluidic device comprising:
• a) at least one tunneling nanotube formation chamber;
• b) a cell transport network of channels connected to at least one inlet and at least one outlet and the at least one tunneling nanotube formation chamber, wherein the cell transport network of channels is a reservoir for cells and cell culture medium and controls the flow rate of the cells and the culture medium through the device, and
• c) a constriction comprising one or more microchannels having a diameter distinct from the diameter of the channels of the cell transport network;
• and
• co-culturing the one or more immature oocyte with the one or more transfected tunneling nanotube-forming cell in the microfluidic device in a manner such that at least one tunneling nanotube-forming cell forms at least one tunneling nanotube to at least one immature oocyte and transfers biomolecules, cellular components, and/or organelles to the at least one immature oocyte through the tunneling nanotube protruded from said at least one tunneling nanotube-forming cell to said at least one immature oocyte for a period of at least one hour.