Establishment of stable cell lines of
 Drosophila
 germ-line stem cells

Niki, Yuzo; Yamaguchi, Takafumi; Mahowald, Anthony P.

doi:10.1073/pnas.0607435103

Cited by 128 publications

(148 citation statements)

References 42 publications

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“…Taken together, these data support a follicular or prefollicular origin of OSS cells, as inferred earlier from their expression of FasIII (Niki et al 2006). …”

Section: ) (Data Not Shown)supporting

confidence: 64%

“…Culture media for OSS cells was prepared as described (Niki et al 2006). For immunostaining, cells were cultured on glass chamber slides coated with Concanavalin-A (Sigma) and fixed in 13 PBS with 4% paraformaldehyde.…”

Section: Oss Cell Methodsmentioning

confidence: 99%

“…For these reasons, identification of homogenous cell lines that express piRNAs would be desirable. Niki et al (2006) described D. melanogaster ovarian cell cultures, including a mixed female germline stem cell/ovarian somatic sheet line (fGS/OSS) and an apparently homogenous derivative culture lacking the germline stem cells termed OSS. We analyzed the OSS line in detail and found that in addition to miRNAs, it expresses high levels of endo-siRNAs and primary piRNAs.…”

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confidence: 99%

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Abundant primary piRNAs, endo-siRNAs, and microRNAs in a Drosophila ovary cell line

et al. 2009

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Piwi proteins, a subclass of Argonaute-family proteins, carry ;24-30-nt Piwi-interacting RNAs (piRNAs) that mediate gonadal defense against transposable elements (TEs). We analyzed the Drosophila ovary somatic sheet (OSS) cell line and found that it expresses miRNAs, endogenous small interfering RNAs (endo-siRNAs), and piRNAs in abundance. In contrast to intact gonads, which contain mixtures of germline and somatic cell types that express different Piwi-class proteins, OSS cells are a homogenous somatic cell population that expresses only PIWI and primary piRNAs. Detailed examination of its TE-derived piRNAs and endo-siRNAs revealed aspects of TE defense that do not rely upon ping-pong amplification. In particular, we provide evidence that a subset of piRNA master clusters, including flamenco, are specifically expressed in OSS and ovarian follicle cells. These data indicate that the restriction of certain TEs in somatic gonadal cells is largely mediated by a primary piRNA pathway.

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“…Taken together, these data support a follicular or prefollicular origin of OSS cells, as inferred earlier from their expression of FasIII (Niki et al 2006). …”

Section: ) (Data Not Shown)supporting

confidence: 64%

Section: Oss Cell Methodsmentioning

confidence: 99%

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confidence: 99%

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Abundant primary piRNAs, endo-siRNAs, and microRNAs in a Drosophila ovary cell line

et al. 2009

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“…GSC-specific genes have been identified using purified GSCs and microarrays [30], while genetic suppressor screens help identify new genes that are important for stem cell regulation [31,32]. It has recently been shown that Drosophila GSCs can be cultured in vitro, together with somatically derived cells, possibly generated by SSCs [33]. In summary, the Drosophila ovary will continue to provide one of the most effective systems for studying stem cell regulation.…”

Section: The Drosophila Ovary Offers Unique Opportunities For Studyinmentioning

confidence: 99%

The Drosophila ovary: an active stem cell community

2007

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Only a small number of cells in adult tissues (the stem cells) possess the ability to self-renew at every cell division, while producing differentiating daughter cells to maintain tissue homeostasis for an organism's lifetime. The Drosophila ovary harbors three different types of stem cell populations (germline stem cell (GSC), somatic stem cell (SSC) and escort stem cell (ESC)) located in a simple anatomical structure known as germarium, rendering it one of the best model systems for studying stem cell biology due to reliable stem cell identification and available sophisticated genetic tools for manipulating gene functions. Particularly, the niche for the GSC is among the first and best studied ones, and studies on the GSC and its niche have made many unique contributions to a better understanding of relationships between stem cells and their niche. So far, both the GSC and the SSC have been shown to be regulated by extrinsic factors originating from their niche and intrinsic factors functioning within. Multiple signaling pathways are required for controlling GSC and SSC self-renewal and differentiation, which provide unique opportunities to investigate how multiple signals from the niche are interpreted in the stem cell. Since the Drosophila ovary contains three types of stem cells, it also provides outstanding opportunities to study how multiple stem cells in a given tissue work collaboratively to contribute to tissue function and maintenance. This review highlights recent major advances in studying Drosophila ovarian stem cells and also discusses future directions and challenges.

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“…To date, several protocols for ex vivo cultivation of Drosophila organs and tissues including testes and germ-line cysts have been developed (e.g., see 4,14,17,20,39,40 ). The cultivation medium and the general conditions used in the protocol presented here were established in 1979 17 .…”

Section: Discussionmentioning

confidence: 99%

Ex vivo Culture of Drosophila Pupal Testis and Single Male Germ-line Cysts: Dissection, Imaging, and Pharmacological Treatment

Gärtner

Rathke

Renkawitz‐Pohl

et al. 2014

JoVE

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During spermatogenesis in mammals and in Drosophila melanogaster, male germ cells develop in a series of essential developmental processes. This includes differentiation from a stem cell population, mitotic amplification, and meiosis. In addition, post-meiotic germ cells undergo a dramatic morphological reshaping process as well as a global epigenetic reconfiguration of the germ line chromatin-the histone-toprotamine switch.Studying the role of a protein in post-meiotic spermatogenesis using mutagenesis or other genetic tools is often impeded by essential embryonic, pre-meiotic, or meiotic functions of the protein under investigation. The post-meiotic phenotype of a mutant of such a protein could be obscured through an earlier developmental block, or the interpretation of the phenotype could be complicated. The model organism Drosophila melanogaster offers a bypass to this problem: intact testes and even cysts of germ cells dissected from early pupae are able to develop ex vivo in culture medium. Making use of such cultures allows microscopic imaging of living germ cells in testes and of germ-line cysts. Importantly, the cultivated testes and germ cells also become accessible to pharmacological inhibitors, thereby permitting manipulation of enzymatic functions during spermatogenesis, including post-meiotic stages.The protocol presented describes how to dissect and cultivate pupal testes and germ-line cysts. Information on the development of pupal testes and culture conditions are provided alongside microscope imaging data of live testes and germ-line cysts in culture. We also describe a pharmacological assay to study post-meiotic spermatogenesis, exemplified by an assay targeting the histone-to-protamine switch using the histone acetyltransferase inhibitor anacardic acid. In principle, this cultivation method could be adapted to address many other research questions in pre-and post-meiotic spermatogenesis.

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Establishment of stable cell lines of Drosophila germ-line stem cells

Cited by 128 publications

References 42 publications

Abundant primary piRNAs, endo-siRNAs, and microRNAs in a Drosophila ovary cell line

Abundant primary piRNAs, endo-siRNAs, and microRNAs in a Drosophila ovary cell line

The Drosophila ovary: an active stem cell community

<em>Ex vivo</em> Culture of <em>Drosophila</em> Pupal Testis and Single Male Germ-line Cysts: Dissection, Imaging, and Pharmacological Treatment

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