Purification of avian circulating primordial germ cells by Nycodenz density gradient centrifugation

Zhao, Dongfeng; Kuwana, Takashi

doi:10.1080/0007166031000085382

Cited by 67 publications

(53 citation statements)

References 17 publications

(19 reference statements)

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“…Circulating PGCs can be recovered from embryonic blood at stages 13-15 (Hamburger and Hamilton, 1951) and concentrated by Ficoll density gradient centrifugation (Yasuda et al, 1992), filtration (Tajima et al, 2003) or Nycodenz density gradient centrifugation (Zhao et al, 2003).…”

Section: Conservation Of Avian Genetic Resources Through Germline Chimentioning

confidence: 99%

Conservation of Avian Genetic Resources

Tajima

2013

J. Poult. Sci.

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Conservation of avian genetic resources is important, as 13.0% (1,313/10,064) of known avian species are currently categorized as endangered by the International Union for Conservation of Nature and Natural Resources (IUCN, 2012). Development of an integrated system for conservation of avian genetics is necessary; however, germplasm conservation systems developed for mammals cannot be applied directly to avians, primarily due to the unique anatomical and physiological characteristics of the avian egg. Thus an alternative strategy for conservation of oviparous species of animals must be developed. Recent technological developments for producing germline chimeras by the transfer of primordial germ cells into recipient embryos has enabled the conservation and retrieval of chicken genetic resources in their complete form. Furthermore, the possibility of propagating endangered wild birds through interspecies germline chimeras has been proposed.In the present review, the current status of avian genetic resource conservation is presented with a focus on the following 4 topics: 1) semen cryopreservation, 2) germline chimeras, 3) somatic nuclear transfer, and 4) other related technologies.

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Section: Conservation Of Avian Genetic Resources Through Germline Chimentioning

confidence: 99%

Conservation of Avian Genetic Resources

Tajima

2013

J. Poult. Sci.

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“…Enrichment of the initial PGC populations generally is used by the Nycodenz density gradient centrifugation method (Zhao and Kuwana 2003) or a Ficoll density gradient (Park et al 2003). Collected PGCs were washed and dispersed in KAv-1 medium (Kuwana et al 1996).…”

Section: Isolation Of Pgcs and Generation Of Pgc-derived Progenymentioning

confidence: 99%

Primordial germ cells (PGCs) as a tool for creating transgenic chickens

Chojnacka-Puchta¹,

Kasperczyk²,

Płucienniczak³

et al. 2012

Polish Journal of Veterinary Sciences

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The transgenic chicken has great potential as a bioreactor for the production of valuable pharmaceutical proteins, notably in the oviduct/egg. Whereas conventional transgenic approaches have significant limitations in this species, an alternative approach employing primordial germ cells (PGCs), the progenitor cells to ova and spermatozoa, has now been successfully applied to the insertion of exogenous genes into birds. Recent developments in manipulating avian embryos make it possible to produce germline chimeras derived from transferred PGCs. In this review we describe the migration pathway of chicken PGCs during early development. We then summarize different methods for the isolation of PGCs and the diversity of techniques used to introduce genes into these cells. Finally, we describe an in vitro assay for testing tissue-specific vectors designed to express heterologous proteins in transgenic chickens.

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“…본 연구와 동일한 발생 단계인 생식선 유래의 gonadal PGC(gPGC)를 이용한 연구 (Natio et al, 1994;Natio, 2003;Zhao and Kuwana, 2003;Mozdziak et al, 2005) (Fig. 3A) 그리고 한국육종협회3호종 (Fig.…”

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The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye)

Kim¹,

Cho²,

Han³

et al. 2014

Korean Journal of Poultry Science

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Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5∼6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid (LN 2 ) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

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Purification of avian circulating primordial germ cells by Nycodenz density gradient centrifugation

Cited by 67 publications

References 17 publications

Conservation of Avian Genetic Resources

Conservation of Avian Genetic Resources

Primordial germ cells (PGCs) as a tool for creating transgenic chickens

The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye)

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