2016
Multivariate Calibration Approach for Quantitative Determination of Cell-Line Cross Contamination by Intact Cell Mass Spectrometry and Artificial Neural Networks
VALLETTA, Elisa; Lukáš KUČERA; Lubomír PROKEŠ; Filippo AMATO; Tiziana PIVETTA et al.Základní údaje
Originální název
Multivariate Calibration Approach for Quantitative Determination of Cell-Line Cross Contamination by Intact Cell Mass Spectrometry and Artificial Neural Networks
Autoři
VALLETTA, Elisa; Lukáš KUČERA; Lubomír PROKEŠ; Filippo AMATO; Tiziana PIVETTA; Aleš HAMPL; Josef HAVEL a Petr VAŇHARA
Vydání
Plos One, San Francisco, Public Library of Science, 2016, 1932-6203
Další údaje
Jazyk
angličtina
Typ výsledku
Článek v odborném periodiku
Obor
Genetika a molekulární biologie
Stát vydavatele
Spojené státy
Utajení
není předmětem státního či obchodního tajemství
Impakt faktor
Impact factor: 2.806
Kód RIV
RIV/00216224:14110/16:00089350
Organizační jednotka
Lékařská fakulta
UT WoS
000369528400026
EID Scopus
2-s2.0-84958825099
Klíčová slova anglicky
EMBRYONIC STEM-CELLS; LASER-DESORPTION/IONIZATION-TIME; LEAST-SQUARES REGRESSION; MALDI-TOF; EXPERIMENTAL-DESIGN; CANCER-DIAGNOSIS; MAMMALIAN-CELLS; CLASSIFICATION; SPECTRA; CULTURE
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 5. 12. 2016 16:59, Ing. Mgr. Věra Pospíšilíková
Anotace
V originále
Cross-contamination of eukaryotic cell lines used in biomedical research represents a highly relevant problem. Analysis of repetitive DNA sequences, such as Short Tandem Repeats (STR), or Simple Sequence Repeats (SSR), is a widely accepted, simple, and commercially available technique to authenticate cell lines. However, it provides only qualitative information that depends on the extent of reference databases for interpretation. In this work, we developed and validated a rapid and routinely applicable method for evaluation of cell culture cross-contamination levels based on mass spectrometric fingerprints of intact mammalian cells coupled with artificial neural networks (ANNs). We used human embryonic stem cells (hESCs) contaminated by either mouse embryonic stem cells (mESCs) or mouse embryonic fibroblasts (MEFs) as a model. We determined the contamination level using a mass spectra database of known calibration mixtures that served as training input for an ANN. The ANN was then capable of correct quantification of the level of contamination of hESCs by mESCs or MEFs. We demonstrate that MS analysis, when linked to proper mathematical instruments, is a tangible tool for unraveling and quantifying heterogeneity in cell cultures. The analysis is applicable in routine scenarios for cell authentication and/or cell phenotyping in general.
Návaznosti
| EE2.3.20.0185, projekt VaV |
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| MUNI/A/1558/2014, interní kód MU |
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| MUNI/M/0041/2013, interní kód MU |
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| ROZV/20/LF/2015, interní kód MU |
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