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

Chojnacka-Puchta, Luiza; Kasperczyk, K.; Płucienniczak, Grażyna; Sawicka, Dorota; Bednarczyk, Marek

doi:10.2478/v10181-011-0132-6

Cited by 22 publications

(16 citation statements)

References 29 publications

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“…Avian oviduct epithelial cells serve as a good model for studying the expression of exogenes delivered by DNA carriers in order to induce the production of human therapeutic proteins in egg white of avian bioreactors (LEE et al 2013;CHOJNACKA-PUCHTA et al 2012;JUNG et al 2011;KAMIHIRA et al 2009). Some results might be extrapolated to human studies related to the hormonal regulation of reproductive tract and diseases caused by pathogens such as avian influenza virus or Salmonella.…”

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confidence: 99%

Efficient Source of Cells in Proximal Oviduct for Testing Non-Viral Expression Constructs in the Chicken Bioreactor Model and for Other in Vitro Studies

Stadnicka

Bodnar

Marszałek

et al. 2016

folia biol (krakow)

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Efficient source of cells in proximal oviduct for testing non-viral expression constructs in the chicken bioreactor model and for other in vitro studies. Folia Biologica (Kraków) 64: 37-46. This work shows the usefulness of chicken oviduct epithelial cells (COEC) in evaluating the efficacy of non-viral expression vectors carrying human therapeutic genes. Secondly, an efficient source of progenitor COEC for in vitro studies is described. Within the distal part of the oviduct, weak to moderate expression of a trans membrane glycoprotein (CD44) was observed. Single cells presenting only weak expression of CD44 were found in magnum sections. in vitro cultured oviduct cells originating from the distal oviduct were suitable for subculturing and showed a stable proliferation potential up to the 2nd passage. However, the pavimentous epithelial-like morphology of COEC was progressively lost over time and mainly a fibroblast-like monolayer was established in consecutive passages. Moreover, various commercial transfection agents including FuGENE6 and XtremeGENE9 DNA were used to optimize delivery of human interferon alfa-2a, (IFNá2a) a therapeutic protein gene under an ovalbumin promoter. The transfection efficiency of adherent COEC was estimated for up to 40% at a ratio of 6:1 of transfectant to pOVA5EIFN + GFP plasmid. Expression of IFNá2a was confirmed by western blotting in transformed COEC. In conclusion, the population of epithelial progenitor cells sourced from the distal oviduct can significantly contribute to in vitro culture of COEC, representing an efficient model to develop the production of avian bioreactors and other in vitro studies related to oviduct tissue.

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confidence: 99%

Efficient Source of Cells in Proximal Oviduct for Testing Non-Viral Expression Constructs in the Chicken Bioreactor Model and for Other in Vitro Studies

Stadnicka

Bodnar

Marszałek

et al. 2016

folia biol (krakow)

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“…In fact, restoration of endangered avian species using SSCs enables the application of chicken germ line transmission technologies to the pheasant, making it possible to produce interspecies germ line chimeras in a chicken-pheasant system. Transfer of foreign genes into chicken embryos, establishment of in vitro culture of germ cells, and production of chimeric chickens and interspecies germ line chimeras [12,13,14,15,16] can be used for the restoration of endangered birds. We attempted to develop interspecies germ cell transfer techniques and to produce interspecies germ line chimeras for the restoration of endangered birds by using SSCs.…”

Section: Introductionmentioning

confidence: 99%

Establishment of a pheasant (Phasianus colchicus) spermatogonial stem cell line for the production of interspecies germ line chimeras

Kim¹,

Sharma²,

Kim³

et al. 2014

Electronic Journal of Biotechnology

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a b s t r a c tBackground: Spermatogonial stem cells (SSCs) are important for the production of interspecies germ line chimeras. The interspecies germ cell transfer technique has been suggested as a way to conserve endangered birds. Our objective was to develop a technique for restoring endangered birds by developing interspecies germ line chimeras between pheasant (Phasianus colchicus) and chicken (Gallus gallus) with SSCs. Results: SSCs were isolated from the surgically removed testis of a pheasant. Growth conditions for pheasant SSCs were established by co-culturing STO (SIM mouse embryo-derived thioguanine and ouabain resistant) cells and pheasant SSCs. The colony-forming cells divided and proliferated stably to yield an established SSC line. Pheasant SSCs showed strong reactivity for GDNF family receptor alpha1 (GFRα1) marker. Finally, production of germ line chimeras was attempted by transferring pheasant SSCs into recipient embryos. Although final embryo survival was 5.6% (20/354), the initial survival rate was 88% (312/354). To measure the percent transfer of donor SSC to gonads, the pheasant SSCs were labeled with PKH 26 fluorescent dye. We observed 30% donor cells and 9.48% c-kit/CD117-positive cells in the gonads of recipient chickens. Donor SSCs were thus stably engrafted in the recipient gonads.Conclusions: This study showed that SSCs can be used as a tool for the conservation of endangered birds and the production of germ line chimeras. Our findings yield insights into how we may use the pheasant spermatogonial stem cell line for efficient production of interspecies germ line chimeras and ultimately, to the restoration of endangered birds.

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“…The primordial germ on poultry has a significant potential to be used in cell-based studies since it can be collected from the blastoderm, embryo blood and embryo gonad (Chojnacka-Puchta et al 2012). Although there is no phenotypic change between the various PGC sources observed and the expression pattern of the certain marker, most PGC can be collected only from one embryo which is an advantage of the gonad PGC (Park & Han 2012).…”

Section: Introductionmentioning

confidence: 99%

Primordial germ cells profiles incubated for 24 hours in phosphate buffer saline [-] solution

Kostaman¹,

Sopiyana²

2018

JITV

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Gonadal development is a sequential process that can be divided into three major events: the PGCs migration, sex determination and gonadal differentiation. This study was aimed to see the development of PGCs isolated from the gonads of embryos after being incubated for 7 days and then was incubated using a solution of Phosphate Buffer Saline (PBS) [-]. The developing gonad can be isolated from 7 days old chick and can be incubated at a temperature of 37.8oC in a solution of PBS [-]: without Ca2+ and Mg2+. The release of gonadal PGC was observed within 1, 8, 16, and 24 hours after the embryonic gonad was placed in a PBS solution [-]. The results showed that PGCs can be separated from gonadal tissues and can be collected by entering the developing gonad to the PBS [-] solution. The highest percentage of PGCs and survival rate was obtained after gonad was incubated for 1 hour and was not different with 8 hours (P>0.05). Those result was significantly different (P<0.05) with the 16 and 24 hours incubation. The highest purity rate percentage was in the 8 hours incubation, but did not show a significant difference (P>0.05) with the 1 and 16 hours incubation. The percentage of the purity differed (P<0.05) after the 24 hours incubation. It can be concluded that the most appropriate incubation time to obtain PGCs from the KUB chicken embryonic gonad is no more than 8 hours.

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

Cited by 22 publications

References 29 publications

Efficient Source of Cells in Proximal Oviduct for Testing Non-Viral Expression Constructs in the Chicken Bioreactor Model and for Other <I>in Vitro</I> Studies

Efficient Source of Cells in Proximal Oviduct for Testing Non-Viral Expression Constructs in the Chicken Bioreactor Model and for Other <I>in Vitro</I> Studies

Establishment of a pheasant (Phasianus colchicus) spermatogonial stem cell line for the production of interspecies germ line chimeras

Primordial germ cells profiles incubated for 24 hours in phosphate buffer saline [-] solution

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