Enhancement of chicken primordial germ cell in vitro maintenance using an automated cell image analyser

Anand, Mahek and Lazar, Bence and Tóth, Roland Imre and Pall, Emoke and Patakiné Várkonyi, Eszter and Liptói, Krisztina and Homolya, László and Hegyi, Zoltán and Gócza, Elen (2018) Enhancement of chicken primordial germ cell in vitro maintenance using an automated cell image analyser. ACTA VETERINARIA HUNGARICA, 66 (4). pp. 518-529. ISSN 0236-6290

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Abstract

Primordial germ cells (PGCs) were isolated from blood samples of chicken embryos. We established four PGC lines: two males (FS-ZZ-101, GFP-ZZ-4ZP) and two females (FS-ZW-111, GFP-ZW-5ZP). We could not detect a significant difference in the marker expression profile, but there was a remarkable difference between the proliferation rates of these PGC lines. We monitored the number of PGCs throughout a three-day period using a high-content screening cell imaging and analysing system (HCS). We compared three different initial cell concentrations in the wells: similar to 1000 cells (1x), similar to 4000 (4x) and similar to 8000 (8x). For the GFP-ZW-5ZP, FS-ZZ-101 and FS-ZW-111 PGC lines the lowest doubling time was observed at 4x concentration, while for GFP-ZZ-4ZP we found the lowest doubling time at 1x concentration. At 8x initial concentration, the growth rate was high during the first two days for all cell lines, but this was followed by the appearance of cell aggregates decreasing the cell growth rate. We could conclude that the difference in proliferation rate could mainly be attributed to genotypic variation in the established PGC lines, but external factors such as cell concentration and quality of the culture medium also affect the growth rate of PGCs.

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