Achieving Optimal Transfection Conditions in Chicken Primordial Germ Cells Under Feeder- and Serum-Free Medium.

ABSTRACT: The successful application of primordial germ cells (PGCs) is an ideal method for generating gene-edited birds. However, barriers to efficient DNA transfection in PGCs lead to low transfection efficiency, limiting the generation of genetically modified chickens. The current study utilized chemical transfection and electroporation methods to determine the optimal transfection conditions for the PGC line under feeder- and serum-free medium. Among the tested methods, the Lonza electroporation system exhibited the highest transduction efficiency, with a previously unreported rate of 71.13 ± 1.26%. Optimal transfection conditions were achieved using 4 µg of DNA and 100 µL of Entranster^TM-E in 1 × 10⁶ PGCs. Furthermore, the optimal electroporation conditions resulted in low

SUBMITTER: Zhao Z 

PROVIDER: S-EPMC11851890 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature