Using Zeocin selection for H9 human ESCs?

14 August 2023 1 5K Report

I am currently establishing a transgenic cell line (H9) using a plasmid containing Zeocin resistant gene. An antibiotic kill curve was performed ranging from 0.5 ug/mL to 1000 ug/mL. H9 WT cells were killed within 2 days with the lowest concentration (0.5 and 1.0 ug/mL), while all cells detached and underwent cell death within a day with other concentration. Similar rate of cell death was observed in cells containing the resistant gene where its gene expression is driven by SV40 promoter.

Therefore, I am asking if anyone has used Zeocin to maintain or select cells with Zeocin antibiotic. If yes, what is the concentration used, particularly on H9 cell lines.

Thank you.

Lei Chen

The use of Zeocin resistance genes is relatively limited. It is necessary to set a gradient concentration to treat cells, and it is generally advisable for all cells to die within about 10 days. Suggest checking other literature or consulting technical personnel.

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