A method for in vitro culture of mammalian ovary tissue based on dynamic regulation of gas-liquid interface

Abstract

The present application relates to the technical field of reproductive biology, and specifically discloses a method for culturing mammalian ovary tissue in vitro based on dynamic regulation of gas-liquid interface. In view of the problem that oxygen supply and nutrient supply cannot be optimized and dynamically regulated in the existing in vitro culture of ovary, the method comprises the following steps: preparing an agar block with culture holes; controlling the liquid level of the culture solution to be flush with the outer edge of the agar block and not higher than the upper surface, so that the culture holes are initially liquid-free; placing the mammalian ovary in the holes and directly exposing it to the gas phase environment in the initial stage; during the culture process, the culture solution gradually enters the holes through capillary action, so that the ovary automatically changes to a semi-submerged state, and dynamic changes of the gas-liquid interface are realized. The present application does not require feeder cells or complex scaffolds, and is simple to operate. Through the synergistic effect of initial gas phase exposure and subsequent semi-submerged state, the internal hypoxia of the ovary is significantly improved, cell apoptosis is reduced, and follicle assembly is promoted, so the present application can be used for follicle development mechanism research and reproductive toxicity evaluation.

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