Enzymatic and Chromosomal Characterization of Hela Variants

Bottomley, R. H.; Trainer, AH; Griffin, Michael K.

doi:10.1083/jcb.41.3.806

Cited by 65 publications

(19 citation statements)

References 14 publications

(13 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The number of chromosomes has been shown to vary between 56 and 179 30, 31, 32, 33. We found the number of kinetochore pairs per cell to be in the range of 38–96 (Fig 1C), in agreement with previous studies.…”

Section: Resultssupporting

confidence: 92%

See 1 more Smart Citation

PRC 1‐labeled microtubule bundles and kinetochore pairs show one‐to‐one association in metaphase

et al. 2016

View full text Add to dashboard Cite

In the mitotic spindle, kinetochore microtubules form k‐fibers, whereas overlap or interpolar microtubules form antiparallel arrays containing the cross‐linker protein regulator of cytokinesis 1 (PRC1). We have recently shown that an overlap bundle, termed bridging fiber, links outermost sister k‐fibers. However, the relationship between overlap bundles and k‐fibers throughout the spindle remained unknown. Here, we show that in a metaphase spindle more than 90% of overlap bundles act as a bridge between sister k‐fibers. We found that the number of PRC1‐GFP‐labeled bundles per spindle is nearly the same as the number of kinetochore pairs. Live‐cell imaging revealed that kinetochore movement in the equatorial plane of the spindle is highly correlated with the movement of the coupled PRC1‐GFP‐labeled fiber, whereas the correlation with other fibers decreases with increasing distance. Analysis of endogenous PRC1 localization confirmed the results obtained with PRC1‐GFP. PRC1 knockdown reduced the bridging fiber thickness and interkinetochore distance throughout the spindle, suggesting a function of PRC1 in bridging microtubule organization and force balance in the metaphase spindle.

show abstract

Section: Resultssupporting

confidence: 92%

“…In HeLa cells, both the number of chromosomes and their structure show abnormalities [27][28][29]. The number of chromosomes has been shown to vary between 56 and 179 [30][31][32][33]. We found the number of kinetochore pairs per cell to be in the range of 38-96 ( Fig 1C), in agreement with previous studies.…”

Section: Resultssupporting

confidence: 91%

PRC 1‐labeled microtubule bundles and kinetochore pairs show one‐to‐one association in metaphase

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Especially for HeLa cells, a wide range of chromosome numbers has been recorded for different strains (e.g. BOTTOMLEY et al 1969;MILLER et al 1971).…”

Section: Chromosome Number and Total Relative Lengthmentioning

confidence: 99%

HeLa cells and their possible contamination of other cell lines: Karyotype studies

Heneen¹

2009

Hereditas

View full text Add to dashboard Cite

G‐ and C‐banding techniques were applied to the chromosomes of 5 human cell lines, viz. HeLa, KB, HEp‐2, D98/AH‐2 and Lu106. According to the records, the 3 former ones are of tumor origin. the 2 latter of normal origin. The 5 cell lines showed considerable similarities: (1) their stemline chromosome numbers were near‐triploid; (2) the normal chromosome types. generally present in disomic or trisomic numbers, showed the same deviations through 4 or all 5 lines: nulli‐ or monosomy in No. 3, tri‐ or tetrasomy in No. 7, 15 and 17, maximum variation in No. 7, 9 and 16, no Y chromosome in spite of the supposedly male origin of some of the lines; (3) the incidence of markers was of comparable size order (21–25% in KB, HeLa and D98/AH‐2, 29% in HEp‐2, 36% in Lu106); (4) the majority of the marker types were found either in relatively low or high frequencies (1–3 or 8–10 out of 10 cells); (5) marker chromosomes of concordant band pattern were common to different lines; (6) the total lengths of normal and marker chromosomes and whole complements were approximately the same in most or all lines. These results indicate that the variability in the parameters analyzed is less than expected in completely independent lines. The conclusion is that all 5 cell lines are most likely of HeLa origin.

show abstract

“…Second, HeLa cells have been reported for decades to 74 contaminate and dominate other cell lines 23,24 and conversely, they are less likely to be 75 contaminated themselves, minimizing the possibility of cell misidentification for HeLa-focused 76 studies 15,16 . Third, due to extensive genome instability during passaging and between labs HeLa 77 cells were reported to contain an exceptional number of genomic variants 18,[25][26][27][28] . For example, the 78 currently widely used HeLa variants include HeLa CCL2, the "original" HeLa cell line; HeLa S3 79 (also called CCL2.2), the third clone isolated from an early HeLa culture; and HeLa Kyoto, the 80 popular Kyoto version.…”

Section: Introductionmentioning

confidence: 99%

Genomic, Proteomic and Phenotypic Heterogeneity in HeLa Cells across Laboratories: Implications for Reproducibility of Research Results

Liu

Mueller

et al. 2018

Preprint

View full text Add to dashboard Cite

1The independent reproduction of research results is a cornerstone of experimental research, yet it is 2 beset by numerous challenges, including the quality and veracity of reagents and materials. Much of 3 life science research depends on life materials, including human tissue culture cells. In this study we 4 aimed at determining the degree of variability in the molecular makeup and the ensuing phenotypic 5 consequences in commonly used human tissue culture cells. We collected 14 stock HeLa aliquots 6 from 13 different laboratories across the globe, cultured them in uniform conditions and profiled the 7 genome-wide copy numbers, mRNAs, proteins and protein turnover rates via genomic techniques 8 and SWATH mass spectrometry, respectively. We also phenotyped each cell line with respect to the 9 ability of transfected Let7 mimics to modulate Salmonella infection. 10 11We discovered significant heterogeneity between HeLa variants, especially between lines of the 12 CCL2 and Kyoto variety. We also observed progressive divergence within a specific cell line over 13 50 successive passages. From the aggregate multi-omic datasets we quantified the response of the 14 cells to genomic variability across the transcriptome and proteome. We discovered organelle-specific 15 proteome remodeling and buffering of protein abundance by protein complex stoichiometry, 16 mediated by the adaptation of protein turnover rates. By associating quantitative proteotype and 17 phenotype measurements we identified protein patterns that explained the varying response of the 18 different cell lines to Salmonella infection. 19 20Altogether the results indicate a striking degree of genomic variability, the rapid evolution of 21 genomic variability in culture and its complex translation into distinctive expressed molecular and 22 phenotypic patterns. The results have broad implications for the interpretation and reproducibility of 23 research results obtained from HeLa cells and provide important basis for a general discussion of the 24 value and requirements for communicating research results obtained from human tissue culture cells. 25 7 molecular analyses, we centrally cultured all 14 cell lines for an additional three passages in our 126 group. To minimize experimental variability, the same researcher cultured the cells using the same 127 culture media and protocol. 128 129As a starting point for the evaluation of HeLa cell heterogeneity, we measured gene copy number 130 variation (CNV) by array-CGH (aCGH). Previous studies reported genome instability within and 131 between different HeLa strains 18, 25-28 . To investigate and display the ploidy variation of the strains 132 studied we projected the CNV data onto the reference human genomic map (Figure 1b). We 133 discovered pervasive CNV differences organized by domains, large chromosomal segments, and 134 even whole chromosomes. Particularly notable were ploidy changes at Chromosomes (Chr) 1, 2, 6, 135 9, 10, 17, 19, 21, 22, and X. (Figure 1b). On average, the HeLa genome of all cells tested...

show abstract

Enzymatic and Chromosomal Characterization of Hela Variants

Cited by 65 publications

References 14 publications

PRC 1‐labeled microtubule bundles and kinetochore pairs show one‐to‐one association in metaphase

PRC 1‐labeled microtubule bundles and kinetochore pairs show one‐to‐one association in metaphase

HeLa cells and their possible contamination of other cell lines: Karyotype studies

Genomic, Proteomic and Phenotypic Heterogeneity in HeLa Cells across Laboratories: Implications for Reproducibility of Research Results

Contact Info

Product

Resources

About