title: Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms.
abstract: Parts of the natural life cycle of Echinococcus granulosus can be retraced in vitro such as the development of protoscoleces into semiadult worms with three or more proglottids, or the redifferentiation of in vitro cultured protoscoleces into metacestode-like cystic structures. Most in vitro generated samples share-at the microscopical level-high similarities with those naturally grown, but developmental differences have also been documented, such as missing egg production in in vitro grown adults or unusual bladder/vesicle formation in protoscoleces cultured into the metacestode direction. The aim of the present study was to explore how far different in vitro generated stage-specific materials/structures match the natural situation on the transcriptome level, based on testing five exemplarily chosen different genes: the frizzled receptor eg-fz4 (posterior marker), the FGF receptor-like factor eg-fgfrl (anterior association), the cell differentiation protein eg-rcd1 (part of the CCR4-NOT complex, a key regulator of eukaryotic gene expression), the rapidly accelerated fibrosarcoma serin/threonin kinase eg-braf (part of the MAPK pathway involved, e.g., in EGF signaling) and the co-smad eg-smadD (downstream factor of TGFβ/BMP2/activin signaling). These genes-tested via qPCR-were selected such as to allow a discussion on their potential role in the development of E. granulosus into the adult stage. Thus, testing took place with three ex vivo isolated samples, namely (i) egg-containing adult worms, (ii) invaginated protoscoleces, and (iii) protoscolex-free germinal layer tissue. Respective data were compared (a) with in vitro generated metacestode-like microcysts developed from protoscolices, and (b) different development stages of protoscoleces in vitro cultured toward adult maturation. As a finding, only eg-smadD and partially eg-fz4 showed high expression similarities between ex vivo harvested and in vitro cultured E. granulosus, thus suggesting a putative role in adult maturation. Conclusively, the fact of using "only" five genes did not allow answering the question if ex vivo and in vitro materials are similar on the transcriptome level. Another experimental restriction arises from different growth conditions of the in vitro cultured materials, and comparing these to the ex vivo harvested ones. Future experiments may solve the problems by using fully standardized E. granulosus sample collection and fully standardized culture conditions.
authors: Research Center for Hydatid Disease in Iran, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.|Institute of Parasitology, Faculty of Medicine and Vetsuisse Faculty of the University of Bern, Bern, Switzerland.
authors: Ebrahim Saedi
authors: ES
authors: Dezaki
authors: Research Department of Biotechnology, Institute of Sciences and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Iran.
authors: Mohammad Mehdi
authors: MM
authors: Yaghoubi
authors: Institute of Parasitology, Faculty of Medicine and Vetsuisse Faculty of the University of Bern, Bern, Switzerland.
authors: Markus
authors: M
authors: Spiliotis
authors: Institute of Parasitology, Faculty of Medicine and Vetsuisse Faculty of the University of Bern, Bern, Switzerland.
authors: Ghalia
authors: G
authors: Boubaker
authors: Department of Pathology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors: Elham
authors: E
authors: Taheri
authors: Research Center for Hydatid Disease in Iran, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors: Pooya Ghaseminejad
authors: PG
authors: Almani
authors: Research Center for Hydatid Disease in Iran, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
authors: Farideh
authors: F
authors: Tohidi
authors: Research Center for Hydatid Disease in Iran, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran. fasihi@kmu.ac.ir.
authors: Majid Fasihi
authors: MF
authors: Harandi
authors: Institute of Parasitology, Faculty of Medicine and Vetsuisse Faculty of the University of Bern, Bern, Switzerland. bruno.gottstein@vetsuisse.unibe.ch.
authors: Bruno
authors: http://orcid.org/0000-0003-0782-3723
authors: B
authors: Gottstein
mesh_terms: D000818:Animals; D004283:Dog Diseases / Q000469:parasitology; D004285:Dogs; D004443:Echinococcosis / Q000469:parasitology / Q000662:veterinary; D004446:Echinococcus; D048209:Echinococcus granulosus / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D020869:Gene Expression Profiling; D017238:Genes, Helminth; D008018:Life Cycle Stages; D008297:Male; D012756:Sheep; D012757:Sheep Diseases / Q000469:parasitology
keywords: Cystic echinococcosis; Developmental genes; Echinococcus granulosus; In vitro cultivation; Real-time qPCR