Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Apr 24;16(5):481.
doi: 10.3390/genes16050481.

Functional Equivalence of Insulin and IGF-1 in the In Vitro Culture of Chicken Primordial Germ Cells

Xin Liu  1   2 , Jun Wu  1   2 , Yixiu Peng  1   2 , Guangzheng Liu  1   2 , Kai Jin  1   2 , Yingjie Niu  1   2 , Jiuzhou Song  3 , Wei Han  4 , Guohong Chen  1   2 , Bichun Li  1   2   5 , Qisheng Zuo  1   2
Affiliations

Functional Equivalence of Insulin and IGF-1 in the In Vitro Culture of Chicken Primordial Germ Cells

Xin Liu et al. Genes (Basel). .

Abstract

Background: Chicken Primordial Germ Cells (PGCs) are one of the few germ cells that can be cultured for a long time in vitro, but challenges remain such as low culture efficiency and unclear roles of nutrient factors and signaling pathways.

Method: In this study, protein kinase B (AKT) pathway activator insulin-like growth factor 1 (IGF-1) was screened for its ability to promote cell proliferation by transcriptome results using various inhibitors of pathway activation. The effects of IGF-1 on PGCs were evaluated through EdU assays, qRT-PCR, flow cytometry, and migration experiments.

Results: This study systematically examined the effects of insulin and IGF-1 on the proliferation, cell cycle, ferroptosis, migration capacity, and establishment efficiency of PGCs. The findings demonstrated that IGF-1 exhibited comparable effects to insulin and could effectively replace insulin in PGC culture systems.

Conclusions: The research results are expected to provide a solid theoretical basis for optimizing the chicken PGC cultivation system and promoting its practical application.

Keywords: IGF-1; chicken PGCs; establishment efficiency; migration.

PubMed Disclaimer

Conflict of interest statement

The authors declare that this research was conducted in the absence of commercial or financial relationships that could be construed as potential conflicts of interest.

Figures

Figure 1
Figure 1
(A) Effects of different activators and inhibitors on the morphology of PGCs; scale bar = 100 μm. (B) Cell count statistics results. Data are expressed as the mean ± SD of 3 independent experiments. Values within a column followed by different superscript letters differ significantly (p < 0.05).
Figure 2
Figure 2
(A) Effects of different activators on the morphology of PGCs; scale bar = 100 μm. (B) Cell count statistics results. Data are expressed as the mean ± SD of 3 independent experiments. Values within a column followed by different superscript letters differ significantly (p < 0.05).
Figure 3
Figure 3
(A) Effects of IGF-1 on morphological observation of PGCs; scale bar = 100 μm. (B,C) Effect of IGF-1 on PGC proliferation. (D,E) EdU proliferation detection and statistical results. Data are expressed as the mean ± SD of 3 independent experiments. Values within a column followed by different superscript letters differ significantly (p < 0.05).
Figure 4
Figure 4
(A,B) Cell cycle detection and statistical analysis were performed using flow cytometry; (C) mRNA expression analysis of cell cycle-related genes, with statistical significance between groups indicated by different letters. Data are expressed as the mean ± SD of 3 independent experiments. Values within a column followed by different superscript letters differ significantly (p < 0.05).
Figure 5
Figure 5
The effect of IGF-1 and insulin media on PI3K/AKT genes in PGCs, with statistical significance between groups indicated by different letters. Data are expressed as the mean ± SD of 3 independent experiments. Values within a column followed by different superscript letters differ significantly (p < 0.05).
Figure 6
Figure 6
(A–D) Flow cytometry analysis of apoptosis and statistical analysis of live, early apoptotic, and late apoptotic cell rates. (E–J) mRNA relative expression analysis of apoptosis-related genes by IGF-1. Different letters in the figure indicate significant differences between groups (p < 0.05).
Figure 7
Figure 7
(A,B) Flow cytometric analysis of the effect of IGF-1 on ROS and fluorescence intensity statistics; (C) statistical analysis of GSH/GSSG content; (D,E) the effect of IGF-1 on MDA and Fe2+ content; (F) the effect and relative expression analysis of IGF-1 on the mRNA expression of ferroptosis-related genes ACSL6 and TFRC. The significant differences between groups are indicated by different letters in the figure (p < 0.05).
Figure 8
Figure 8
Effects of IGF-1 and insulin treatment on the expression and migration ability of key genes in chicken PGCs. (A) Migration images of GFP-PGCs from insulin-free controls, IGF-1, and insulin to the gonads of recipient chicken embryos; (B) PGC migration rate in insulin-free controls, IGF-1, and insulin; (C) the effects of IGF-1 and insulin on pluripotency and marker genes. The significant differences between groups are indicated by different letters in the figure (p < 0.05), NS: not significant.
Figure 9
Figure 9
Effects of IGF-1 and insulin on the efficiency of establishing PGCs cell lines. (A) Representative images at different time points during the establishment of chicken PGCs showing the changes in cell proliferation; (B,C) statistical results of IGF-1 and insulin system establishment efficiency. NS: not significant.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

  • Chicken Primordial Germ Cell Surface Marker.
    Gough TJ, Wise TG, Bruce MP, Doran TJ, Layton DS, Bean AGD. Gough TJ, et al. Animals (Basel). 2025 Jun 24;15(13):1868. doi: 10.3390/ani15131868. Animals (Basel). 2025. PMID: 40646767 Free PMC article.

References

    1. Farzaneh M., Khoshnam S.E., Nokhbatolfoghahai M. First scientific record of two cases of partial twinning in the chick embryo, Gallus gallus domesticus. Vet. Rec. Case Rep. 2016;4:e000353. doi: 10.1136/vetreccr-2016-000353. - DOI
    1. Trefil P., Aumann D., Koslová A., Mucksová J., Beneová B., Kalina J., Wurmser C., Fries R., Elleder D., Schusser B. Male fertility restored by transplanting primordial germ cells into testes: A new way towards efficient transgenesis in chicken. Sci. Rep. 2017;7:14246. doi: 10.1038/s41598-017-14475-w. - DOI - PMC - PubMed
    1. Han J.Y., Park Y.H. Primordial germ cell-mediated transgenesis and genome editing in birds. J. Anim. Sci. Biotechnol. 2018;9:19. doi: 10.1186/s40104-018-0234-4. - DOI - PMC - PubMed
    1. Woodcock M.E., Gheyas A.A., Mason A.S., Nandi S., Mcgrew M.J. Reviving rare chicken breeds using genetically engineered sterility in surrogate host birds. Proc. Natl. Acad. Sci. USA. 2019;116:201906316. doi: 10.1073/pnas.1906316116. - DOI - PMC - PubMed
    1. Whyte J., Glover J.D., Woodcock M., Brzeszczynska J., Taylor L., Sherman A., Kaiser P., Mcgrew M.J. FGF, Insulin, and SMAD Signaling Cooperate for Avian Primordial Germ Cell Self-Renewal. Stem Cell Rep. 2015;5:1171–1182. doi: 10.1016/j.stemcr.2015.10.008. - DOI - PMC - PubMed

LinkOut - more resources