Establishment and application of coilia ectenes gonad somatic cell line
A technology of somatic cells and scorpionfish, applied in the biological field, can solve the problems of complex cell components and restricting the mechanism of fish spermatogenesis, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0038] Embodiment 1 Cultus sex gland cell culture
[0039] 1. Isolation and Digestion of Gonad Cells from Anchovy
[0040] Take the anchovy from the Yangtze River, soak the fish body in PBS containing 0.5% bleaching powder for 1 min, wash with PBS containing 1% double antibody, pH=7.4 for 3 times, and then dissect. Cut the belly of the fish with scissors, carefully clip out the gonads with tweezers, place them in a petri dish containing 1% double-antibody PBS, carefully peel off the fat tissue on the surface of the gonads with tweezers, wash 3 times with PBS containing 1% double-antibody, and transfer the gonads Transfer to a 1.5mL EP tube containing 500μL of trypsin, cut the tissue properly with scissors, and place it on ice for digestion. After 1 h, 1 mL of medium was added to stop the digestion, and the tissue pieces were gently blown away with a pipette gun, and the cell suspension was transferred to a 24-well plate covered with 0.1% gelatin, and cultured at 28°C.
[004...
Embodiment 2
[0044] The identification of embodiment 2 anchovy sex gland cells
[0045] 1. RT-PCR identification of cultured anchovy sex gland cell types
[0046] In order to identify the type of gonad cells of the cultured Anchovy anchovy, the RNA of the cells was extracted and reversed to cDNA for RT-PCR detection.
[0047] Primer Premier 6 was used to design the primers of sox9, fshr and clu for the sex gland marker genes of Anchovy anchovy and the primers of vasa, dazl and piwi for the germ cell marker genes (see Table 1 for the sequence information of the primers), extract the cellular RNA and convert it into cDNA, and use RT -PCR identification of the gonad cell types of the anchovy.
[0048] Table 1 RT-PCR primer sequence information
[0049] gene name Primer sequence (F: upstream, R: downstream) sox9b F: TGGACCCCTACCTGAAGATG; R: AGTCCAGTCGTAGCCCTTGA fshr F: GTGGTGCTGGTGTTGCTGCTTA; R: TGGACGAGTGAGTAGATAGTGCCTTC clu F: TCTCTGCTCTGTGTCTTATC; R: AACTT...
Embodiment 3
[0055] Example 3 Induction of spermatogonial stem cells from the anchovy sex gland cells
[0056] 1. Cultivation of Spermia Stem Cells
[0057] A. Isolation and Digestion of Testicular Cells from Snormouth Fish
[0058] Take juvenile horse mouth fish, soak the fish body in PBS containing 0.5% bleaching powder for 1 min, wash with PBS containing 1% double antibody for 3 times, and then dissect. Cut the belly of the fish with scissors, carefully clip out the testis with tweezers, place it in a petri dish containing 1% double-antibody PBS, carefully peel off the fat tissue on the surface of the testis with tweezers, wash 3 times with PBS containing 1% double-antibody, and transfer the testis Transfer to a 1.5mL EP tube containing 500μL of trypsin, cut the tissue properly with scissors, and place it on ice for digestion. After 1 h, 1 mL of medium was added to stop the digestion, and the tissue pieces were gently blown away with a pipette gun, and the cell suspension was transfer...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com