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Direct in vitro propagation of avian germ cells from an embryonic gonad biorepository

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Author(s):
Hu, Tuanjun ; Purdy, Phillip H. ; Blank, Marcel H. ; Muhonja, Christine K. ; Pereira, Ricardo J. G. ; Tiambo, Christian K. ; McGrew, Mike J.
Total Authors: 7
Document type: Journal article
Source: POULTRY SCIENCE; v. 103, n. 11, p. 9-pg., 2024-09-07.
Abstract

Direct introduction of cryopreserved embryonic gonadal germ cells (GGC) into a sterile chicken surrogate host to reconstitute a chicken breed has been demonstrated as a feasible approach for preserving and utilizing chicken genetic resources. This method is highly efficient using male gonads; however, a large number of frozen female embryonic gonads is needed to provide sufficient purified GGC for the generation of fertile surrogate female hosts. Applying this method to indigenous chicken breeds and other bird species is difficult due to small flock numbers and poor egg production in each egg laying cycle. Propagating germ cells from the frozen gonadal tissues may be a solution for the biobanking of these birds. Here, we describe a simplified method for culture of GGC from frozen embryonic 9.5 d gonads. At this developmental stage, the germ cells are autonomously shed into medium, yielding hundreds to thousands of mitosis-competent germ cells. The resulting cultures of GGC have over 90% purity, uniformly express SSEA-1 and DAZL antigens and can re-colonize recipient's gonads. The GGC recovery rate from frozen gonads are 42% to 100%, depending on length of cryopreservation and the breed or line of chickens. Entire chicken embryos can also be directly cryopreserved for later gonadal isolation and culture. This storage method is a supplementary approach to safeguard local indigenous chicken breeds bearing valuable genetic traits and should be applicable to the biobanking of many bird species. (AU)

FAPESP's process: 22/03780-6 - Interspecific transplant of iPSCs cells using genetically engineered sterile host: a model for conservation of endangered birds of Pernambuco Endemism Center
Grantee:Marcel Henrique Blank
Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
FAPESP's process: 20/11479-9 - Generation of induced pluripotent stem cell (iPSC) in endangered wild animals
Grantee:Marcel Henrique Blank
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 16/17158-4 - CONSERVATION OF ENDANGERED CURASSOWS THROUGH INTERSPECIFIC GERM CELL TRANSPLANTATION
Grantee:Ricardo Jose Garcia Pereira
Support Opportunities: Regular Research Grants
FAPESP's process: 17/23548-2 - Evaluation, recovering and conservation of endangered animal species from the Pernambuco Centre of Endemism
Grantee:Luís Fábio Silveira
Support Opportunities: BIOTA-FAPESP Program - Thematic Grants