Corneal endothelial cell culture method

• Main Authors: ZHAN ZHENJIAN, CAI QIUYU, ZHENG SHIXIN, LI JUN, DONG WENJUAN, LIU SHIHAO, SU YUNFEI, LI YAORAO, LI XUBAI, QI GUOGUANG
• Format: Patent
• Language: Chinese; English
• Published: 13.10.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention provides a culture method of corneal endothelial cells, which comprises the following steps: culturing unfrozen corneal endothelial cells by using a resuscitation culture solution in a resuscitation stage, and culturing by using a basic culture solution containing coriolus versicolor polysaccharide in a passage stage to obtain target corneal endothelial cells, the resuscitation culture solution is prepared by adding 200 to 300 mL/L of fetal calf serum, 2 to 3 [mu] g/mL of recombinant insulin, 15 to 18 ng/mL of endothelial cell growth factors, 25 to 30 ng/mL of basic fibroblast growth factors, 5 to 8 [mu] g/mL of lutein ester, 2 to 4 [mu] g/mL of dunaliella salina flavine, 0.5 to 0.8 mM of adenine and 5 to 8 ng/mL of gentamicin into a DMEM-F12 culture solution. The method has a certain influence on the adherent growth of the corneal endothelial cells, so that when the corneal endothelial cells resuscitated by freezing are cultured in vitro for 4 hours, the cell adherent rate reaches 70% or above,