Migration and Proliferation of Intact and Genetically Modified Primordial Germ Cells and the Generation of a Transgenic Chicken1

Abstract

This study evaluated gonadal migration and postmigratory proliferation of intact and genetically modified chicken primordial germ cells (PGCs). A randomized, controlled trial was conducted with the gonadal population of PGCs and transgenic chicken production as major parameters. PGCs (0, 90, 900, 1800, or 3000 cells) were transferred into 53-h-old embryos. The percentage of PGCs migrating on Day 6 of development was highest (35.8%) following the transfer of 900 PGCs and did not change with increases in transferred PGCs. The number of migrating PGCs gradually increased (P = 0.0001) as the number of transferred PGCs was increased. Gonadal migration was detected after the transfer of intact and genetically modified PGCs, but prominent decreases in PGC migration (from 21.9% to 0.38%) and chimera ratio (from 0.4 to 0.007) occurred with genetically modified PGCs. However, subsequent vigorous proliferation of the modified PGCs (3.67-fold increase from transferred number) led to the derivation of a germline chimera and produced a transgenic hatchling. In conclusion, the number of migrating PGCs increased as the number of transferred cells increased. Vigorous proliferation after transfer compensated for the decreased migration capacity of genetically modified PGCs and resulted in the production of a transgenic chicken.