Viability and Functionality of Primordial Germ Cells after Freeze-thaw in Chickens

Nakamura, Yatsuka; Usui, Fumitake; Miyahara, Daichi; Mori, Takafumi; Watanabe, Haruhi; Ono, Tamao; Takeda, Kumiko; Nirasawa, Keijiro; Kono, Hiroshi; Tagami, Takahiro

doi:10.2141/jpsa.010085

Cited by 19 publications

(29 citation statements)

References 28 publications

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“…Commercially available serum and DMSO-based cryomedium also results in higher recovery and viability of chicken PGCs after thawing than a common freezing protocol [55]. Due to its easier availability and higher performance, commercial cryomedium particularly CELLBANKER 1 (Nippon Zenyaku Kogyo, Koriyama, Japan) has been used to cryopreserve PGCs from chicken and quail [30, 59,60,61]. In contrast to the slow-freezing method, a vitrification method can theoretically store cells both intracellularly and extracellularly by ice-free solidification.…”

Section: Ultra-low Temperature Preservation Of Pgcsmentioning

confidence: 99%

Poultry genetic resource conservation using primordial germ cells

Nakamura

2016

Journal of Reproduction and Development

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The majority of poultry genetic resources are maintained in situ in living populations. However, in situ conservation of poultry genetic resources always carries the risk of loss owing to pathogen outbreaks, genetic problems, breeding cessation, or natural disasters. Cryobanking of germplasm in birds has been limited to the use of semen, preventing conservation of the W chromosome and mitochondrial DNA. A further challenge is posed by the structure of avian eggs, which restricts the cryopreservation of ova and fertilized embryos, a technique widely used for mammalian species. By using a unique biological property and accessibility of avian primordial germ cells (PGCs), precursor cells for gametes, which temporally circulate in the vasculature during early development, an avian PGC transplantation technique has been established. To date, several techniques for PGC manipulation including purification, cryopreservation, depletion, and long-term culture have been developed in chickens. PGC transplantation combined with recent advanced PGC manipulation techniques have enabled ex situ conservation of poultry genetic resources in their complete form. Here, the updated technologies for avian PGC manipulation are introduced, and then the concept of a poultry PGC-bank is proposed by considering the biological properties of avian PGCs.

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Section: Ultra-low Temperature Preservation Of Pgcsmentioning

confidence: 99%

Poultry genetic resource conservation using primordial germ cells

Nakamura

2016

Journal of Reproduction and Development

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“…2A, B). Naito et al (1994) and Nakamura et al (2011) were reported that PGCs propagation abilities in vivo are varied between chicken breeds, and in vitro proliferative of PGC-LCs also might be different between WL and BPR.…”

Section: Discussionmentioning

confidence: 99%

Culture Conditions for Maintain Propagation, Long-term Survival and Germline Transmission of Chicken Primordial Germ Cell-Like Cells

et al. 2014

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Transplantation of primordial germ cells (PGCs), which are the progenitor cells of gametes, is a powerful tool for generation of transgenic chickens. However, the frequencies of transgene integration into the genome of purified PGCs still remain low. An in vitro culture system enabling chicken PGCs to propagate efficiently would be useful for efficient transgenesis of PGCs. In the present study, we optimized the culture conditions for chicken PGCs to enhance the proliferation and evaluated the germline transmission of cultured PGCs that proliferated for long periods of time. PGC-like cells (PGC-LCs), that have remarkably similar morphological characteristics to intact PGCs, could be derived by cultivation of blood containing PGCs obtained from 2.5-day-old chicken embryos according to the protocol of van de Lavoir et al. (2006). We determined which feeder cells and which growth factors were required to improve proliferation of PGC-LCs. Male PGC-LCs survival and proliferation were enhanced during culture in the basic medium containing either basic fibroblast growth factor (bFGF) alone or both bFGF and stem cell factor (SCF) on a feeder of buffalo rat liver (BRL) cells. Male PGC-LCs could be propagated in defined culture condition for extended periods. These cells expressed the germline-specific protein Vasa and undifferentiated cell marker stage-specific embryonic antigen-1 (SSEA-1) and pluripotency genes Nanog and PouV. Furthermore, Male PGC-LCs cultured for 225 d could migrate toward and colonize within recipient gonads and transmit to the next generation following transplantation. We succeeded in produce 3 offspring originating from long-term cultured PGC-LCs from a germline chimeric rooster (6%). The present study represents valuable steps toward defining a culture condition enabling PGCLCs to propagate efficiently for long periods in vitro with maintenance of their commitment to the germline.

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“…It has recently been indicated that the proliferative ability of PGCs after settlement in recipient gonads is different between chicken breeds (Nakamura et al . ). When the same number of cPGCs from three different chicken breeds is transferred into the bloodstream of recipient embryo, the number of exogenous PGCs that settle in the gonads of a 6‐day‐old (stage 29) recipient is significantly different between the three breeds.…”

Section: Germ Cell Competition In Chimeric Gonadsmentioning

confidence: 97%

Development, differentiation and manipulation of chicken germ cells

2013

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Germ cells are the only cell type capable of transmitting genetic information to the next generation. During development, they are set aside from all somatic cells of the embryo. In many species, germ cells form at the fringe of the embryo proper and then traverse through several developing somatic tissues on their migration to the emerging gonads. Primordial germ cells (PGCs) are the only cells in developing embryos with the potential to transmit genetic information to the next generation. Unlike other species, in avian embryos, PGCs use blood circulation for transport to the future gonadal region. This unique accessibility of avian PGCs during early development provides an opportunity to collect and transplant PGCs. The recent development of methods for production of germline chimeras by transfer of PGCs, and long-term cultivation methods of chicken PGCs without losing their germline transmission ability have provided important breakthroughs for the preservation of germplasm , for the production of transgenic birds and study the germ cell system. This review will describe the development, migration, differentiation and manipulation of germ cells, and discuss the prospects that germ cell technologies offer for agriculture, biotechnology and academic research.

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Viability and Functionality of Primordial Germ Cells after Freeze-thaw in Chickens

Cited by 19 publications

References 28 publications

Poultry genetic resource conservation using primordial germ cells

Poultry genetic resource conservation using primordial germ cells

Culture Conditions for Maintain Propagation, Long-term Survival and Germline Transmission of Chicken Primordial Germ Cell-Like Cells

Development, differentiation and manipulation of chicken germ cells

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