Derivation of embryonic stem cells across avian species

Abstract

Copyright and License

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Data Availability

The RNA-seq datasets generated in this study have been uploaded to the Gene Expression Omnibus under the accession numbers GSE254284 (chicken) and GSE298550 (quail, goose and ostrich). Publicly available RNA-seq datasets used in this study: mouse ES cells in naive, formative and primed states. GSE131555 (ref. ⁶⁴); chicken embryos from EGK.I to EGK.VIII. GSE86592 (ref. ⁸⁶); chicken embryos at HH4. SRX540095 (Weizmann Institute). Publicly available reference genomes used in this study: chicken reference genome (Ensembl GRCg6_106), quail reference genome (Ensembl Coturnix_japonica.Coturnix_japonica_2.0.113), goose reference genome (Ensembl Anser_cygnoides.GooseV1.0.113) and Ostrich reference genome (Ensembl Struthio_camelus_australis.ASM69896v1.113).

Acknowledgement

We thank the Proteomics Core Facility at the University of Southern California for liquid chromatography with tandem mass-spectrometry analysis; the Millard and Muriel Jacobs Genetics and Genomics Laboratory at the California Institute of Technology for next-generation sequencing; the Molecular Cytogenetics Laboratory at Texas A&M University for karyotype analysis; Miyazaki City Phoenix Zoo (Miyazaki, Japan), Kumamoto City Zoological and Botanical Gardens (Kumamoto), Kurume Bird Center (Fukuoka), Hirakawa Zoological Park (Kagoshima), Osaki Peacock Park (Nagasaki) and Nansei Environmental Laboratory Co., Ltd (Okinawa) for their generous supply of fertilized blue peafowl eggs; D. McKim for valuable comments, discussions and proofreading of the paper; and M. Nakakubo, N. Matsubara and T. Seto for their technical assistance. Funding: this work was supported by the Chen Yong Foundation of the Zhongmei Group, the Wu & Jiang Research Fund and the Xia Research Fund: grant nos. NIH/NIGMS R01 GM151373 and NIH/NIGMS R41 GM146516 to Q.-L.Y. and the Revive & Restore Biotechnology for Bird Conservation grants to Q.-L.Y., G.S. and C.L. C.-M.C. and P.W. were supported by grant nos. NIH/NIAMS R01 AR047364 and AR060306 and the research contract no. 005884 between the University of Southern California (USC) and the China Medical University in Taiwan. G.S. was supported by Japan Society for the Promotion of Science grant nos. 18H02452 and 21H02490, the Japan Science and Technology Agency e-ASIA Joint Research Project no. JPMJSC19E5 and the Takeda Science Foundation Research Grant. X.C. was supported by the predoctoral fellowship from the NICHD/USC Joint T32 Training Program in Developmental Biology, Stem Cells and Regeneration, and a Della Martin postdoctoral Fellowship in Mental Illness at Caltech. Z.G. was supported by the California Institute for Regenerative Medicine Predoctoral Training Fellowship in Stem Cell Biology and Tissue Regeneration.

Conflict of Interest

Q.-L.Y., X.C., Z.G., X.T. and X.L. are inventors on a patent (WO2023158627A3) entitled ‘Methods for derivation and propagation of avian pluripotent stem cells and applications thereof’ arising from this work. The other authors declare no competing interests.

Supplemental Material

Supplementary Table 2

Supplementary Table 3

Supplementary Table 4

Supplementary Table 5

Primer sequences used in this study.

Supplementary Video 1

Supplementary Video 2

Contribution of GFP-labeled chicken ES cells to the beating heart of a nonirradiated E4 chicken embryo. Constitutively GFP-expressing chicken ES cells were injected into the subgerminal cavity of a nonirradiated EGK.X-stage chicken embryo and incubated using the surrogate shell system. On day 4, live imaging of green fluorescence was performed.

Supplementary Video 3

Supplementary Video 4

GFP-labeled donor chicken ES cells contribute to blood vessels and circulating cells in a sublethally irradiated E4 chicken embryo. This video features the same embryo shown in Supplementary Video 3. Constitutively GFP-expressing chicken ES cells were injected into the subgerminal cavity of an EGK.X-stage embryo treated with 500 cGy and incubated using the surrogate shell system. On day 4, live imaging of green fluorescence in the extra-embryonic region was recorded.

Supplementary Video 5

GFP-labeled donor quail ES cells contribute to circulating cells in a sublethally irradiated E4 chicken embryo. Constitutively GFP-expressing quail ES cells were injected into the subgerminal cavity of an EGK.X-stage chicken embryo treated with 500 cGy. The embryo was incubated using the surrogate shell system. On day 4, live imaging of green fluorescence in the extra-embryonic region was recorded.

Supplementary Video 6

GFP-labeled donor goose ES cells contribute to blood vessels and circulating cells in a sublethally irradiated E5 chicken embryo. Constitutively GFP-expressing goose ES cells were injected into the subgerminal cavity of an EGK.X-stage chicken embryo treated with 500 cGy. The embryo was incubated using the surrogate shell system. On day 5, live imaging of green fluorescence in the extra-embryonic region was recorded.

Supplementary Video 7