Project 5: Cell isolation
Cell isolation and establishment of cell lines from Schistosoma mansoni
One of several limitations in current schistosome research is the lack of schistosome cell lines. These would serve as a continuous source for proteins, RNA, and DNA. Furthermore, such cell lines would facilitate the functional analysis of schistosome genes, or the recombinant expression of schistosomal proteins for immunisation purposes (antibody production) and the identification of putative vaccines1,2. In addition, the establishment of schistosome cell lines could help to reduce animal experiments (3R principle). Recently, S1-S4 vitelline cells have been successfully isolated on the basis of a novel organ isolation method3 and intensively characterised4. Future plans comprise the characterisation of such cells at the molecular level (transcriptomics) and the optimisation of protocols for the isolation of cells from other tissues. In this context one focus will be to investigate cells with stem-cell character such as S1-vitelline cells or oogonia. Furthermore, we aim to establish protocols for primary cell cultures of S. mansoni but also to immortalise cells to obtain permanent cell lines for this parasite.
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| C | D |
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Microscopic analyses of gonadal cells of S. mansoni females. A, bright-field microscopy (BFM) of isolated vitelline cells4. Their differentiation stages (indicated as S1 – S4) can be assigned according to their size and granularity of the cell content. B, scanning electron microscopy (SEM) of vitelline cells of different stages of development (S1 - S4). C, BFM of oocytes from S. mansoni females; D, SEM of oocytes from S. mansoni females. iO: immature oocytes; mO: mature oocytes; scale bars: 10 µM. |
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1Quack T, Wippersteg V, Grevelding CG. (2010) Cell cultures for schistosomes - chances of success or wishful thinking? Int J Parasitol. 40(9):991-1002.
2Ye Q, Dong HF, Grevelding CG, Hu M. (2013) In vitro cultivation of Schistosoma japonicum-parasites and cells. Biotechnol Adv. 31(8):1722-37.
3Hahnel S, Lu Z, Wilson RA, Grevelding CG, Quack T. (2013) Whole-organ isolation approach as a basis for tissue-specific analyses in Schistosoma mansoni. PLoS Negl Trop Dis. 7(7):e2336.
4Lu Z, Quack T, Hahnel S, Gelmedin V, Pouokam E, Diener M, Hardt M, Michel G, Baal N, Hackstein H, Grevelding CG. (2015) Isolation, enrichment and primary characterisation of vitelline cells from Schistosoma mansoni obtained by the organ isolation method. Int J Parasitol. 45(9-10):663-72.