Rapid enrichment and separation method for gonad somatic cells of plectropomus leopardus

By employing flexible fluorescent labeling and single-cell selection techniques, combined with optimized culture media, the problem of low cell survival rate during the isolation of leopard gill spiny perch gonadal cells was solved, achieving efficient and stable cell isolation and culture, and supporting reproductive stem cell research.

CN121825856AActive Publication Date: 2026-04-10HAINAN UNIVERSITY SANYA NANFAN RESEARCH INSTITUTE +1
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Patent Information

Application Number
CN202610316519.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-03-16
Publication Date
2026-04-10
Estimated Expiration
2046-03-16

AI Technical Summary

Technical Problem

Existing technologies struggle to maintain high-precision screening of gonadal cells in leopard-gill perch while ensuring cell survival, leading to poor reproductive development and low reproductive efficiency in the leopard-gill perch aquaculture industry.

Method used

A method combining flexible fluorescent labeling, single-cell mechanical selection, and optimized cell culture conditions was employed, including trypsin-EDTA digestion, polysorbate treatment, fluorescence microscopy verification, and the use of a single-cell micropicking instrument. This method, combined with specific antibodies and optimized culture medium, enabled the rapid enrichment and isolation of gonadal cells.

Benefits of technology

High-purity, high-viability gonadal cells were isolated, exhibiting high cell adhesion and strong proliferation capacity, making them suitable for long-term stable passage. This fills the gap in the in vitro culture system of gonadal cells from the leopard gill spiny perch and supports subsequent experimental research.

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Abstract

The invention relates to the technical field of aquatic animal cell culture, in particular to a rapid enrichment and separation method for gonad somatic cells of plectropomus leopardus. Comprising the following steps: selecting healthy plectropomus leopardus for anesthesia; the method comprises the following steps: taking gonad tissues, performing sterile treatment, cutting, centrifugal washing and filtering, inoculating the gonad tissues into a culture medium for primary culture to obtain gonad cells emigrated from tissue blocks, performing trypsin-EDTA digestion and centrifugal washing, and treating the gonad cells with 1 * PBS containing polysorbate; after closing, antibody incubation and washing, the marking effect is verified through a fluorescence microscope, and specific enrichment of gonad somatic cells is achieved; and filtering and purifying the enriched fluorescence-labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescence-positive cells under a fluorescence visual field, transferring the fluorescence-positive cells into a culture dish containing a complete medium, inoculating the separated fluorescence-positive cells into the optimized complete medium, and carrying out purification culture, so that the cells grow in a wall-attached manner and stable passage is realized.
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