Culture solution for mechanical separation of cytoplasm and use method of culture solution

Abstract

The invention discloses a culture solution for mechanical exfoliation of cytoplasm and a use method thereof, the culture solution comprises in vitro operation culture solution basic components, additive components and the like, and the culture solution is characterized in that the additive components comprise substances for softening cell membranes during mechanical exfoliation of cytoplasm; this includes, but is not limited to, the addition or combined addition of polyvinylpyrrolidone (PVP), cytochalasin, protein components (such as serum or serum proteins or substitutes thereof) at appropriate concentrations. The culture solution is favorable for realizing smooth separation of cytoplasm from cell bodies under the action of mechanical force under the conditions of micromanipulation and the like, that is, when the culture solution is used, ovum, cells or partial cell bodies taken down from the ovum are placed in the culture solution, so that cell membranes become soft and viscous, and when the mechanical force is applied, for example, a microscopic tube is used for sucking one part of the cell body to swing or extrude and the like, the cytoplasm of the stressed part is easily separated from the primitive cell body, the cell membrane of the separated part and the non-separated part is still kept complete, and the cell membrane disintegration is not easy to occur.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture medium for mechanically detaching cytoplasm and a use method thereof. Background technique [0002] At present, the well-known micro-operations such as micro-insemination or nucleoplasm replacement need to be carried out under a microscope, out of the incubator environment with a certain concentration of carbon dioxide, using in vitro operation culture fluid or buffer medium, such as mHTF medium is a synthetic, defined solution for the handling and transfer of human gametes and embryos. This buffer system does not require the use of a CO2 incubator. Its components include: water; salts and ions, such as sodium chloride, potassium chloride, magnesium carbonate (or magnesium sulfate), calcium chloride, etc.; energy substrates, such as sodium pyruvate, sodium lactate, etc.; antibiotics (such as Qing Damycin) and acid-base indicators (such as phenol red), etc.; especially inc...

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Problems solved by technology

[0004] In order to overcome the problem that the existing operating culture solution cannot smoothly carry out the mechanical detachment of the cytoplasm, the present invention makes full use of the new functions of the above-mentioned additives (polyvinylpyrrolidone, cytochalasin B, protein components) discovered by the inventors in practice, namely The softness and viscosity of the cell membrane are used for new operating purposes, and the deficiency of its existing conventional concentration is found, thus establishing a new culture medium and a new set of operating methods

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