Methods in cell science : an official journal of the Society for In Vitro Biology最新文献

{"title":"Establishment of cell culture systems from penaeid shrimp and their susceptibility to white spot disease and yellow head viruses.","authors":"S N Chen,&nbsp;C S Wang","doi":"10.1023/a:1009885929335","DOIUrl":"https://doi.org/10.1023/a:1009885929335","url":null,"abstract":"<p><p>Monolayer cultures were established from ovary, heart, lymphoid tissue and peripheral hemocytes of penaeid shrimps including Penaeus monodon, P. japonicus and P. penicillatus. The most favorable conditions for the culture of penaeid shrimp cells in vitro was in CMRL and L-15 tissue culture media when used within an osmolarity range of 620--760 mmol/kg. The optimal maintenance temperature was 25 degrees C for tissues of P. japonicus and 28 degrees C for tissues of P. monodon and P. penicillatus. Among the four tissues tested, lymphoid tissue, or 'Oka organ', was superior to the other tissues for the formation of confluent cell monolayers. Cell cultures from lymphoid tissue and ovary have been subcultured up to three times. When peripheral hemocytes and heart were cultured, a maximum survival of 4 days was obtained. In contrast, cell cultures derived from ovary and lymphoid tissue were maintained alive for at least 20 days in appropriate culture systems. Neither confluent cell sheet nor adherence of cells was obtained in cultivation of hepatopancreas using the present culture systems. The results obtained from the present study also revealed that ovary extract, muscle extract and lobster hemolymph enhanced the survival of the cultured cells of penaeid shrimp in vitro. When the 'Oka organ' cell monolayer was incubated with either white spot disease virus (WSDV) or yellow head virus (YHV), no cytopathic effect (CPE) was obtained. However, at 5--7 days after establishment, significant CPE (a few foci) was observed in cell monolayers derived from WSDV- and YHV-infected Oka tissue. By electron microscopy, virions of WSDV and YHV were observed in the nuclei and cytoplasm of cultured cells. The CPE foci developed further with increased incubation time.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 4","pages":"199-206"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009885929335","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21482875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Early attempts at production of prawn cell lines.","authors":"L Owens,&nbsp;J Smith","doi":"10.1023/a:1009806606562","DOIUrl":"https://doi.org/10.1023/a:1009806606562","url":null,"abstract":"<p><p>This report describes some unsuccessful attempts to produce continuous cell lines from penaeid prawn tissues in the late 1980s. This information is presented so that others might save time by not repeating the unsuccessful measures that were attempted. The osmolarity of Penaeus monodon haemolymph was measured at 687 mOsmol/kg (N = 10). Of the media tested, the best medium for cell growth and maintenance was shown to be double strength L-15, supplemented with 10% foetal bovine serum, and 10% prawn muscle extract at 28 degrees C ( approximately 675.5 mOsmol/kg). Prawn muscle extract was made by homogenizing 30 g of prawn muscle in 50/50 ratio of distilled water/autoclaved seawater, clarified stepwise by centrifugation at 2k, 14k, 14k xg for 30 minutes each. The resultant supernatant was heat-inactivated on occasions with no improvement in growth. Preconditioned medium, cholesterol, galactose and trehalose supplements and the use of Cell-Tak did not improve growth conditions, and haemolymph extracts were detrimental to the cells. In addition it was shown that Nunc 25 cm(2) plastic culture flasks were better than Linbro and both were better than glass as substrates. The fate of 101 individual primary cell cultures, established from penaeid prawns, was as follows. Fifteen of the cultures succumbed to bacterial contamination, five became contaminated with fungi, four with thauastrochytrids, four succumbed to presumptive viral autocultures and two to ciliate contamination. Cell cultures derived from heart tissue could be maintained for a mean of 12.7 days (sd 9.7d), those derived from the epidermis 15.6 days (sd 9.0d), ovarian tissue 10 days (sd 2d), lymphoid organ 6.8 days (sd 0.4d), nerve cord and hepatopancreas 2 days. The most persistent cell cultures -- those derived from the heart explants -- contained dividing cells at 40 days, and epidermis cells were still dividing at 30 days. The longest lasting, non- proliferating, but viable, cell cultures were those of subcutis/epidermis and heart cells which remained viable for 240 and 307 days respectively. Only cell cultures from multiple prawns achieved 100% confluency in 25 cm(2) plastic culture flasks. No cultures survived attempts at passage by either trypsinisation or mechanical disruption with a rubber policeman. No cell lines were established.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 4","pages":"207-12"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009806606562","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21482876","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synchronization of somatic embryogenesis at high frequency using carrot suspension cultures: model systems and application in plant development.","authors":"K Osuga,&nbsp;H Masuda,&nbsp;A Komamine","doi":"10.1023/a:1009884806166","DOIUrl":"https://doi.org/10.1023/a:1009884806166","url":null,"abstract":"<p><p>Materials and methods for the high frequency induction and synchronous somatic embryogenesis from cultured cells of higher plants are described, using carrot suspension cultures as a model system of higher plants. The following four synchronous systems of somatic embryogenesis, which were established in our laboratories, are reported: (1) Somatic embryogenesis from single cells. a) Small spherical single cells, obtained from suspension cultures in the presence of 2,4-D, zeatin and mannitol by sieving, density gradient centrifugation in Percoll solutions and manual picking up, form embryogenic cell clusters, which differentiate to embryos at high frequency, when embryogenic cell clusters are transferred to a medium lacking 2,4-D. b) Explants of hypocotyls of regenerated plantlets from somatic embryos were cultured after treatment with 2,4-D for 12-24 h, and then transferred into a fresh medium lacking 2,4-D. Single cells are released from hypocotyl explants and differentiated into embryos at high frequency. In this system, a large number of single cells and embryogenic cells can be collected. (2) Somatic embryogenesis from embryogenic cell clusters, which are obtained from suspension cultures by sieving, density gradient centrifugation in Ficoll solutions, and subsequent centrifugation at a low speed, differentiate synchronously to globular embryos at high frequency. Plantlets are formed from globular embryos. (3) Embryogenic cell clusters obtained according to the procedure described in (2) are cultured at cell densities of 2x10(3) cell clusters ml(-1). Globular embryos differentiate to torpedo-shaped embryos and subsequently to plantlets at high frequency when they are cultured at densities below 150 globular embryos ml(-1).</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 2-3","pages":"129-40"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009884806166","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21580175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mutagensis and cell transformation in cell culture.","authors":"M S Crane","doi":"10.1023/a:1009861505170","DOIUrl":"https://doi.org/10.1023/a:1009861505170","url":null,"abstract":"<p><p>The current lack of continuous prawn cell lines suitable for the isolation and growth of prawn viruses is a major setback for diagnosis of viral diseases of prawns; isolation and identification of the causative agents are severely hindered and the development of other diagnostic procedures is slowed. To date, there are few, if any, examples of continuous prawn cell lines suitable for virus isolation and growth. Recent studies indicate that clastogenic agents such as ionising radiation are more effective than powerful point mutagens as immortalizing agents for mammalian cells. Such studies with mammalian cell cultures indicate that certain mutagenic agents and/or prolonged treatment with a combination of mutagens may prove more useful in the production of immortal prawn cell lines in vitro rather than other, more limited and specific procedures. This report provides a brief review of the use of mutagens to induce immortalisation -- the acquisition of unlimited growth potential -- in mammalian cells in culture, i.e. the production of continuous cell lines from primary cultures. Aspects of cell transformation vs. immortalisation, the control of cell growth in vitro, and the application of mutagenic agents to induce immortalisation in primary cultures of prawn tissues will be discussed.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 4","pages":"245-53"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009861505170","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21482882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A method for three-dimensional coculture of cancer cells combined to any other type of cells maintained organotypically.","authors":"R Beaupain","doi":"10.1023/a:1009899527587","DOIUrl":"https://doi.org/10.1023/a:1009899527587","url":null,"abstract":"<p><p>A three-dimensional cell coculture method is presented where cancer cells can be maintained alone or combined with other cell types in longterm culture in order to reconstitute some of the interactions between the different cell elements in tumors in vivo. The cells are accumulated by centrifugation to form 'nodules' which are cultivated on a semisolid agar medium at medium/air interface. The nodules are not mere cell aggregates, they are able to develop morphological and functional differentiation as well as tissue-like membrane junctions. Studies on short-term and long-term effects of anticancer treatments are possible and their long-term regrowth can be obtained. Especially, in nodules containing cell mixtures, the localization of the different cell types can be determined and their specific differentiation. An example showing stroma-like formations and collagen production in breast cancer cell and breast fibroblast containing nodules is presented.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 1","pages":"25-30"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009899527587","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21584805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preparation of primary cell cultures from lamprey.","authors":"C Ma,&nbsp;P Collodi","doi":"10.1023/a:1009824303498","DOIUrl":"https://doi.org/10.1023/a:1009824303498","url":null,"abstract":"<p><p>The lamprey is an important model for studies of evolution and comparative biology. The ability to culture cells from lamprey tissues makes it possible to employ an in vitro approach to address basic questions in these areas. Methods are described for the initiation of cell cultures derived from tissues of adult and larval sea lamprey (Petromyzon marinus). Primary cultures initiated from gill, muscle, gut, brain, ovary, heart and kidney were viable for up to eight months and several of the cultures were propagated for multiple passages. Most cultures were initiated from tissue explants in basal nutrient medium supplemented with fetal bovine and trout sera on a culture surface treated with fibronectin and collagen. Variations of these culture conditions to meet the specific growth requirements of certain cell types are discussed.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 1","pages":"39-46"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009824303498","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21584807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bivariate flow cytometry DNA/BrdUrd analysis of plant cell cycle.","authors":"S Lucretti,&nbsp;L Nardi,&nbsp;P T Nisini,&nbsp;F Moretti,&nbsp;G Gualberti,&nbsp;J Dolezel","doi":"10.1023/a:1009893008892","DOIUrl":"https://doi.org/10.1023/a:1009893008892","url":null,"abstract":"<p><p>We describe a protocol for flow cytometry analysis of cell cycle in plants using indirect immunolabelling staining and Vicia faba, Pisum sativum and Zea mays root tip cells as model systems. The protocol is based on simultaneous analysis of two fluorescent signals. The first, obtained after staining with propidium iodide, is used to quantify nuclear DNA content. The second, obtained after indirect immunofluorescent staining of bromodeoxyuridine (BrdUrd), is used to quantify the amount of BrdUrd incorporated into nuclear DNA. In an attempt to standardize the procedure, the effects of various conditions for partial DNA denaturation using HCl, as well as of BrdUrd concentration and incorporation time on flow cytometry DNA/BrdUrd content analysis have been studied. Maximum BrdUrd-linked fluorescence was observed after a 30 min pulse with 10 microM BrdUrd and after DNA denaturation with 1.5 N HCl (final concentration) for 30 min at 25 degrees C. Under these conditions, DNA content histograms with relatively small coefficient of variation (< 4%, full peak) could be obtained. To avoid non-specific staining of cytoplasm and cell walls, the protocol involves the use of nuclei isolated from formaldehyde-fixed tissues. Fixed isolated nuclei are stable and may be stored in hexylene glycol 0.75 M at 4 degrees C for prolonged periods prior to actual staining and analysis.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 2-3","pages":"155-66"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009893008892","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21580126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Primary cell cultures isolated from Penaeus monodon prawns.","authors":"L West,&nbsp;T Mahony,&nbsp;F McCarthy,&nbsp;J Watanabe,&nbsp;D Hewitt,&nbsp;S Hansford","doi":"10.1023/a:1009864212013","DOIUrl":"https://doi.org/10.1023/a:1009864212013","url":null,"abstract":"<p><p>We have devised a cell culture system for Penaeus monodon prawn cells that uses a defined synthetic medium. Organs were removed from adult prawns ranging in size from 13--19cm rostrum-to-telson length. Cultures consisted of either a blend of hematopoietic and lymphoid cells or ovarian cells. The cells divide rapidly in culture, doubling on average once per week for 5 to 6 weeks. These cultures continue to survive for at least 5 months but the rates of cell division are low after the first 5--6 weeks. In the literature, unicellular eukaryotic marine organisms such as chytrids may contaminate marine cell cultures. In some cases these eukaryotic contaminants may be difficult to distinguish from prawn cells unless detailed ultrastructure or characteristic developmental stages such as zoospores can be observed. Alternatively, we prepared molecular probes from repeated DNA sequences 100--400 bp in length in the P. monodon genome. These species-specific probes were hybridised to genomic DNA from cell culture to confirm proliferation of P. monodon cells in our cultures.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 4","pages":"219-23"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009864212013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21482878","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Crustacean primary cell culture: A technical approach.","authors":"J Y Toullec","doi":"10.1023/a:1009833924792","DOIUrl":"https://doi.org/10.1023/a:1009833924792","url":null,"abstract":"<p><p>Crustacean cell culture has gained attention as a potent model to assist in the development of diagnostic reagents and probes for use in the shrimp, crayfish and lobster industries. The availability of such cellular tools is especially important to industries which use intensive aquaculture methods and thus have increased risk of disease problems. Indeed, crustacean cell cultures offer potential for studying the effects of pathogens in vitro and for increasing our knowledge on developmental and sexual maturation processes, or endocrine regulation in crustaceans. Although numerous attempts have been undertaken, no established cell line of marine crustaceans has been reported to date. However, primary cultures obtained from various organ sources are reported with increasing frequency. They represent the first steps towards the establishment of cell lines and they provide useful information concerning the most suitable cell culture conditions involved in the survival and proliferative capacity of the various tissues.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 4","pages":"193-8"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009833924792","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21482874","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Primary culture of lymphoid organ cells and haemocytes of kuruma shrimp, Penaeus japonicus.","authors":"T Itami,&nbsp;M Maeda,&nbsp;M Kondo,&nbsp;Y Takahashi","doi":"10.1023/a:1009845103353","DOIUrl":"https://doi.org/10.1023/a:1009845103353","url":null,"abstract":"<p><p>A primary cell culture system was developed for the cells of lymphoid organ tissue of kuruma shrimp, Penaeus japonicus. Minced tissues of lymphoid organs were seeded and incubated at 30 degrees C in medium 199 supplemented with 20% foetal bovine serum, a salt mixture and a lactalbumin hydrolysate (0.1 g/l). Fibroblast-like cells and epithelioid-like cells survived for 54 days. Cells did not survive after trypsin, collagenase or hyaluronidase treatment used for cell dissociation. Mitogens (Con A, PHA-P, Pokeweed) and insulin did not enhance cell proliferation. When penaeid rod-shaped DNA virus (PRDV) was inoculated into the lymphoid organ cell culture, a cytopathic effect was observed within 8 days. On the other hand, large granular haemocytes that were fractionated using a Percoll continuous density gradient were not infected with PRDV in vitro within 10 days, which was the longest period of haemocyte maintenance.</p>","PeriodicalId":80082,"journal":{"name":"Methods in cell science : an official journal of the Society for In Vitro Biology","volume":"21 4","pages":"237-44"},"PeriodicalIF":0.0,"publicationDate":"1999-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/a:1009845103353","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21482881","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}