Animal Bioscience

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
권호 표지
DOI QR코드

DOI QR Code

Isolation and characterization of cultured chicken oviduct epithelial cells and in vitro validation of constructed ovalbumin promoter in these cells

  • Yang, Hyeon (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Bo Ram (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Hwi-Cheul (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jung, Sun Keun (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Ji-Youn (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • No, Jingu (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Shanmugam, Sureshkumar (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jo, Yong Jin (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Haesun (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Hwang, Seongsoo (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Byun, Sung June (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration)
  • Received : 2020.09.04
  • Accepted : 2020.12.07
  • Published : 2021.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: Transgenic hens hold a great promise to produce various valuable proteins. Through virus transduction into stage X embryo, the transgene expression under the control of constructed chicken ovalbumin promoters has been successfully achieved. However, a validation system that can evaluate differently developed ovalbumin promoters in in vitro, remains to be developed. Methods: In the present study, chicken oviduct epithelial cells (cOECs) were isolated from oviduct tissue and shortly cultured with keratinocyte complete medium supplemented with chicken serum. The isolated cells were characterized with immunofluorescence, western blot, and flow cytometry using oviduct-specific marker. Chicken mutated ovalbumin promoter (Mut-4.4-kb-pOV) was validated in these cells using luciferase reporter analysis. Results: The isolated cOECs revealed that the oviduct-specific marker, ovalbumin protein, was clearly detected by immunofluorescence, western blot, and flow cytometry analysis revealed that approximately 79.40% of the cells contained this protein. Also, luciferase reporter analysis showed that the constructed Mut-4.4-kb-pOV exhibited 7.1-fold (p<0.001) higher activity in the cOECs. Conclusion: Collectively, these results demonstrate the efficient isolation and characterization of cOECs and validate the activity of the constructed ovalbumin promoter in the cultured cOECs. The in vitro validation of the recombinant promoter activity in cOECs can facilitate the production of efficient transgenic chickens for potential use as bioreactors.

Keywords

Acknowledgement

This work was supported with the National Institute of Animal Science (Grant No. PJ01260301), Rural Development Administration (RDA), Republic of Korea

References

  1. Ivarie R. Competitive bioreactor hens on the horizon. Trends Biotechnol 2006;24:99-101. https://doi.org/10.1016/j.tibtech.2006.01.004
  2. McGrew MJ, Sherman A, Ellard FM, et al. Efficient production of germline transgenic chickens using lentiviral vectors. EMBO Rep 2004;5:728-33. https://doi.org/10.1038/sj.embor.7400171
  3. Lee J, Ma J, Lee K. Direct delivery of adenoviral CRISPR/ Cas9 vector into the blastoderm for generation of targeted gene knockout in quail. Proc Natl Acad Sci USA 2019;116: 13288-92. https://doi.org/10.1073/pnas.1903230116
  4. Part TS, Lee HG, Moon JK, et al. Deposition of bioactive human epidermal growth factor in the egg white of transgenic hens using an oviduct-specific minisynthetic promoter. FASEB J 2015;29:2386-96. https://doi.org/10.1096/fj.14-264739
  5. Cao D, Wu H, Li Q, et al. Expression of recombinant human lysozyme in egg whites of transgenic hens. PLoS One 2015; 10:e0118626. https://doi.org/10.1371/journal.pone.0118626
  6. Liu T, Wu H, Cao D, et al. Oviduct-specific expression of human neutrophil defensin 4 in lentivirally generated transgenic chickens. PLoS One 2015;10:e0127922. https://doi.org/10.1371/journal.pone.0127922
  7. Dougherty DC, Sanders MM. Estrogen action: revitalization of the chick oviduct model. Trends Endocrinol Metab 2005; 16:414-9. https://doi.org/10.1016/j.tem.2005.09.001
  8. Lillico SG, Sherman A, McGrew MJ, et al. Oviduct-specific expression of two therapeutic proteins in transgenic hens. Proc Natl Acad Sci USA 2007;104:1771-6. https://doi.org/10.1073/pnas.0610401104
  9. Kwon SC, Choi JW, Jang H-J, et al. Production of biofunctional recombinant human interleukin 1 receptor antagonist (rhIL1RN) from transgenic quail egg white. Biol Reprod 2010;82:1057-64. https://doi.org/10.1095/biolreprod.109.081687
  10. Herron LR, Pridans C, Turnbull ML, et al. A chicken bioreactor for efficient production of functional cytokines. BMC Biotechnol 2018;18:82. https://doi.org/10.1186/s12896-0180495-1
  11. Sanders MM, McKnight GS. Chicken egg white genes: multihormonal regulation in a primary cell culture system. Endocrinology 1985;116:398-405. https://doi.org/10.1210/endo-116-1-398
  12. Kim SH, Choi JY, Shin C-R, et al. Induction of apoptosis in chicken oviduct cells by C2-ceramide. Mol Cells 2005;19: 185-90.
  13. Jung JG, Park TS, Kim JN, et al. Characterization and application of oviductal epithelial cells in vitro in Gallus domesticus. Biol Reprod 2011;85:798-807. https://doi.org/10.1095/biolreprod.111.092023
  14. Kasperczyk K, Bajek A, Joachimiak R, et al. In vitro optimization of the Gallus domesticus oviduct epithelial cells culture. Theriogenology 2012;77:1834-45. https://doi.org/10.1016/j.theriogenology.2011.12.029
  15. Stadnicka K, Slawinska A, Dunislawska A, Pain B, Bednarczyk M. Molecular signatures of epithelial oviduct cells of a laying hen (Gallus gallus domesticus) and quail (Coturnix japonica). BMC Dev Biol 2018;18:9. https://doi.org/10.1186/s12861018-0168-2
  16. Scott BB, Lois C. Generation of tissue-specific transgenic birds with lentiviral vectors. Proc Natl Acad Sci USA 2005;102: 16443-7. https://doi.org/10.1073/pnas.0508437102
  17. Byun SJ, Kim SW, Kim K-W, et al. Oviduct-specific enhanced green fluorescent protein expression in transgenic chickens. Biosci Biotechnol Biochem 2011;75:646-9. https://doi.org/10.1271/bbb.100721
  18. Dierich A, Gaub MP, LePennec JP, Astinotti D, Chambon P. Cell-specificity of the chicken ovalbumin and conalbumin promoters. EMBO J 1987;6:2305-12. https://doi.org/10.1002/j.1460-2075.1987.tb02505.x
  19. Ebers KL, Zhang CY, Zhang MZ, Bailey RH, Zhang S. Transcriptional profiling avian beta-defensins in chicken oviduct epithelial cells before and after infection with Salmonella enterica serovar Enteritidis. BMC Microbiol 2009;9:153. https://doi.org/10.1186/1471-2180-9-153
  20. Kohler PO, Grimley PM, O'Malley BW. Estrogen-induced cytodifferentiation of the ovalbumin-secreting glands of the chick oviduct. J Cell Biol 1969;40:8-27. https://doi.org/10.1083/jcb.40.1.8
  21. Palmiter RD. Interaction of estrogen, progesterone, and testosterone in the regulation of protein synthesis in chick oviduct. Biochemistry 1971;10:4399-403. https://doi.org/10.1021/bi00800a008
  22. Sevane N, Bialade F, Velasco S, et al. Dietary inulin supplementation modifies significantly the liver transcriptomic profile of broiler chickens. PLoS One 2014;9:e98942. https://doi.org/10.1371/journal.pone.0098942
  23. Couteaudier M, Trapp-Fragnet L, Auger N, et al. Derivation of keratinocytes from chicken embryonic stem cells: establishment and characterization of differentiated proliferative cell populations. Stem Cell Res 2015;14:224-37. https://doi.org/10.1016/j.scr.2015.01.002

Cited by

  1. Development and in vitro evaluation of recombinant chicken promoters to efficiently drive transgene expression in chicken oviduct cells vol.100, pp.10, 2021, https://doi.org/10.1016/j.psj.2021.101365