Dataset Information

Ultra-sensitive detection of tumorigenic cellular impurities in human cell-processed therapeutic products by digital analysis of soft agar colony formation.

ABSTRACT: Contamination with tumorigenic cellular impurities is one of the most pressing concerns for human cell-processed therapeutic products (hCTPs). The soft agar colony formation (SACF) assay, which is a well-known in vitro assay for the detection of malignant transformed cells, is applicable for the quality assessment of hCTPs. Here we established an image-based screening system for the SACF assay using a high-content cell analyzer termed the digital SACF assay. Dual fluorescence staining of formed colonies and the dissolution of soft agar led to accurate detection of transformed cells with the imaging cytometer. Partitioning a cell sample into multiple wells of culture plates enabled digital readout of the presence of colonies and elevated the sensitivity for their detection. In practice, the digital SACF assay detected impurity levels as low as 0.00001% of the hCTPs, i.e. only one HeLa cell contained in 10,000,000 human mesenchymal stem cells, within 30 days. The digital SACF assay saves time, is more sensitive than in vivo tumorigenicity tests, and would be useful for the quality control of hCTPs in the manufacturing process.

SUBMITTER: Kusakawa S

PROVIDER: S-EPMC4672267 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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Ultra-sensitive detection of tumorigenic cellular impurities in human cell-processed therapeutic products by digital analysis of soft agar colony formation.

Kusakawa Shinji S Yasuda Satoshi S Kuroda Takuya T Kawamata Shin S Sato Yoji Y

Scientific reports 20151208

Contamination with tumorigenic cellular impurities is one of the most pressing concerns for human cell-processed therapeutic products (hCTPs). The soft agar colony formation (SACF) assay, which is a well-known in vitro assay for the detection of malignant transformed cells, is applicable for the quality assessment of hCTPs. Here we established an image-based screening system for the SACF assay using a high-content cell analyzer termed the digital SACF assay. Dual fluorescence staining of formed ...[more]

PMID: 26644244

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Project description:BACKGROUND: Production and consumption of industrially processed food and drink products have risen in parallel with the global increase in overweight and obesity and related chronic non-communicable diseases. The objective of this study was to analyze the relationship between household availability of processed and ultra-processed products and the prevalence of excess weight (overweight plus obesity) and obesity in Brazil. METHODS: The study was based on data from the 2008-2009 Household Budget Survey involving a probabilistic sample of 55,970 Brazilian households. The units of study were household aggregates (strata), geographically and socioeconomically homogeneous. Multiple linear regression models were used to assess the relationship between the availability of processed and ultra-processed products and the average of Body Mass Index (BMI) and the percentage of individuals with excess weight and obesity in the strata, controlling for potential confounders (socio-demographic characteristics, percentage of expenditure on eating out of home, and dietary energy other than that provided by processed and ultra-processed products). Predictive values for prevalence of excess weight and obesity were estimated according to quartiles of the household availability of dietary energy from processed and ultra-processed products. RESULTS: The mean contribution of processed and ultra-processed products to total dietary energy availability ranged from 15.4% (lower quartile) to 39.4% (upper quartile). Adjusted linear regression coefficients indicated that household availability of ultra-processed products was positively associated with both the average BMI and the prevalence of excess weight and obesity, whereas processed products were not associated with these outcomes. In addition, people in the upper quartile of household consumption of ultra-processed products, compared with those in the lower quartile, were 37% more likely to be obese. CONCLUSION: Greater household availability of ultra-processed food products in Brazil is positively and independently associated with higher prevalence of excess weight and obesity in all age groups in this cross-sectional study.

Dataset Information

Ultra-sensitive detection of tumorigenic cellular impurities in human cell-processed therapeutic products by digital analysis of soft agar colony formation.

Publications

Ultra-sensitive detection of tumorigenic cellular impurities in human cell-processed therapeutic products by digital analysis of soft agar colony formation.

Similar Datasets

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