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排序方式: 共有75条查询结果,搜索用时 31 毫秒
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Rita Barallon Steven R. Bauer John Butler Amanda Capes-Davis Wilhelm G. Dirks Eugene Elmore Manohar Furtado Margaret C. Kline Arihiro Kohara Georgyi V. Los Roderick A. F. MacLeod John R. W. Masters Mark Nardone Roland M. Nardone Raymond W. Nims Paul J. Price Yvonne A. Reid Jaiprakash Shewale Gregory Sykes Anton F. Steuer Douglas R. Storts Jim Thomson Zenobia Taraporewala Christine Alston-Roberts Liz Kerrigan 《In vitro cellular & developmental biology. Animal》2010,46(9):727-732
Cell misidentification and cross-contamination have plagued biomedical research for as long as cells have been employed as research tools. Examples of misidentified cell lines continue to surface to this day. Efforts to eradicate the problem by raising awareness of the issue and by asking scientists voluntarily to take appropriate actions have not been successful. Unambiguous cell authentication is an essential step in the scientific process and should be an inherent consideration during peer review of papers submitted for publication or during review of grants submitted for funding. In order to facilitate proper identity testing, accurate, reliable, inexpensive, and standardized methods for authentication of cells and cell lines must be made available. To this end, an international team of scientists is, at this time, preparing a consensus standard on the authentication of human cells using short tandem repeat (STR) profiling. This standard, which will be submitted for review and approval as an American National Standard by the American National Standards Institute, will provide investigators guidance on the use of STR profiling for authenticating human cell lines. Such guidance will include methodological detail on the preparation of the DNA sample, the appropriate numbers and types of loci to be evaluated, and the interpretation and quality control of the results. Associated with the standard itself will be the establishment and maintenance of a public STR profile database under the auspices of the National Center for Biotechnology Information. The consensus standard is anticipated to be adopted by granting agencies and scientific journals as appropriate methodology for authenticating human cell lines, stem cells, and tissues. 相似文献
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Nims RW Sykes G Cottrill K Ikonomi P Elmore E 《In vitro cellular & developmental biology. Animal》2010,46(10):811-819
The role of cell authentication in biomedical science has received considerable attention, especially within the past decade.
This quality control attribute is now beginning to be given the emphasis it deserves by granting agencies and by scientific
journals. Short tandem repeat (STR) profiling, one of a few DNA profiling technologies now available, is being proposed for
routine identification (authentication) of human cell lines, stem cells, and tissues. The advantage of this technique over
methods such as isoenzyme analysis, karyotyping, human leukocyte antigen typing, etc., is that STR profiling can establish
identity to the individual level, provided that the appropriate number and types of loci are evaluated. To best employ this
technology, a standardized protocol and a data-driven, quality-controlled, and publically searchable database will be necessary.
This public STR database (currently under development) will enable investigators to rapidly authenticate human-based cultures
to the individual from whom the cells were sourced. Use of similar approaches for non-human animal cells will require developing
other suitable loci sets. While implementing STR analysis on a more routine basis should significantly reduce the frequency
of cell misidentification, additional technologies may be needed as part of an overall authentication paradigm. For instance,
isoenzyme analysis, PCR-based DNA amplification, and sequence-based barcoding methods enable rapid confirmation of a cell
line’s species of origin while screening against cross-contaminations, especially when the cells present are not recognized
by the species-specific STR method. Karyotyping may also be needed as a supporting tool during establishment of an STR database.
Finally, good cell culture practices must always remain a major component of any effort to reduce the frequency of cell misidentification. 相似文献
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Temperature is one of the main factors that determine sexual reproduction in terrestrial and emergent aquatic plant species. The effect of temperature on sexual reproduction and seed production of Glyceria maxima (Hartm.) Holmb. in the southern hemisphere is unknown. Glyceria maxima collections in February 2010 at three isolated infestations in KwaZulu-Natal failed to yield a single seed, only empty panicles. Laboratory experiments showed that vernalisation had no consistent effect on seed production. Field- and laboratory-grown plants produced seeds in the 2010/2011 season, because of having sufficient time at optimum temperatures required for seed production (1 491 and 1 585 hours, respectively), compared to a shorter period (1 352 hours) of suitable temperatures during the 2009/2010 growing season. An inadequate period of optimum temperatures (15–25°C) during seed production resulted in the lack of seeds in the field in the 2009/2010 growing season. This study showed that temperature and duration of exposure thereto during the seed-production period play vital roles in G. maxima sexual reproduction. 相似文献
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John M. Baust Gertrude Case Buehring Lia Campbell Eugene Elmore John W. Harbell Raymond W. Nims Paul Price Yvonne A. Reid Frank Simione 《In vitro cellular & developmental biology. Animal》2017,53(8):669-672
This overview describes a series of articles to provide an unmet need for information on best practices in animal cell culture. The target audience primarily consists of entry-level scientists with minimal experience in cell culture. It also include scientists, journalists, and educators with some experience in cell culture, but in need of a refresher in best practices. The articles will be published in this journal over a six-month period and will emphasize best practices in: (1) media selection; (2) use and evaluation of animal serum as a component of cell culture medium; (3) receipt of new cells into the laboratory; (4) naming cell lines; (5) authenticating cell line identity; (6) detecting and mitigating risk of cell culture contamination; (7) cryopreservation and thawing of cells; and (8) storing and shipping viable cells. 相似文献
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VLJ Whitehall TD Dumenil DM McKeone CE Bond ML Bettington RL Buttenshaw L Bowdler GW Montgomery LF Wockner BA Leggett 《Epigenetics》2014,9(11):1454-1460
The CpG Island Methylator Phenotype (CIMP) is fundamental to an important subset of colorectal cancer; however, its cause is unknown. CIMP is associated with microsatellite instability but is also found in BRAF mutant microsatellite stable cancers that are associated with poor prognosis. The isocitrate dehydrogenase 1 (IDH1) gene causes CIMP in glioma due to an activating mutation that produces the 2-hydroxyglutarate oncometabolite. We therefore examined IDH1 alteration as a potential cause of CIMP in colorectal cancer. The IDH1 mutational hotspot was screened in 86 CIMP-positive and 80 CIMP-negative cancers. The entire coding sequence was examined in 81 CIMP-positive colorectal cancers. Forty-seven cancers varying by CIMP-status and IDH1 mutation status were examined using Illumina 450K DNA methylation microarrays. The R132C IDH1 mutation was detected in 4/166 cancers. All IDH1 mutations were in CIMP cancers that were BRAF mutant and microsatellite stable (4/45, 8.9%). Unsupervised hierarchical cluster analysis identified an IDH1 mutation-like methylation signature in approximately half of the CIMP-positive cancers. IDH1 mutation appears to cause CIMP in a small proportion of BRAF mutant, microsatellite stable colorectal cancers. This study provides a precedent that a single gene mutation may cause CIMP in colorectal cancer, and that this will be associated with a specific epigenetic signature and clinicopathological features. 相似文献