Variation in RNA expression and genomic DNA content acquired during cell culture

Hiorns, Lynne R.; Bradshaw, Tracey D.; Skelton, Lorraine; Yu, Qingnan; Kèlland, Lloyd R.; Leyland‐Jones, Brian

doi:10.1038/sj.bjc.6601405

Cited by 30 publications

(29 citation statements)

References 22 publications

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“…Two HZs on 17q (27.06 - 49.35 Mb and 69.51 - 80.48 Mb) detected in our study were shown to be amplified by others using CGH[15]. Although the HZ from 171.40 to 184.09 Mb on 2q is smaller than the predicted minimum size (26 homologous SNPs in cluster compared to 30 for the minimum size of a non-random zone) it was shown to be amplified in some sub-lines of MCF-7.…”

Section: Discussionsupporting

confidence: 62%

Identification of possible genetic alterations in the breast cancer cell line MCF-7 using high-density SNP genotyping microarray

Wang¹,

Greenawalt²,

Cui³

et al. 2009

J Carcinog

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Context:Cancer cell lines are used extensively in various research. Knowledge of genetic alterations in these lines is important for understanding mechanisms underlying their biology. However, since paired normal tissues are usually unavailable for comparison, precisely determining genetic alterations in cancer cell lines is difficult. To address this issue, a highly efficient and reliable method is developed. Aims: Establishing a highly efficient and reliable experimental system for genetic profiling of cell lines.Materials and Methods:A widely used breast cancer cell line, MCF-7, was genetically profiled with 4,396 single nucleotide polymorphisms (SNPs) spanning 11 whole chromosomes and two other small regions using a newly developed high-throughput multiplex genotyping approach.Results:The fractions of homozygous SNPs in MCF-7 (13.3%) were significantly lower than those in the control cell line and in 24 normal human individuals (25.1% and 27.4%, respectively). Homozygous SNPs in MCF-7 were found in clusters. The sizes of these clusters were significantly larger than the expected based on random allelic combination. Fourteen such regions were found on chromosomes 1p, 1q, 2q, 6q, 13, 15q, 16q, 17q and 18p in MCF-7 and two in the small regions.Conclusions:These results are generally concordant with those obtained using different approaches but are better in defining their chromosomal positions. The used approach provides a reliable way to detecting possible genetic alterations in cancer cell lines without paired normal tissues.

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Section: Discussionsupporting

confidence: 62%

Identification of possible genetic alterations in the breast cancer cell line MCF-7 using high-density SNP genotyping microarray

Wang¹,

Greenawalt²,

Cui³

et al. 2009

J Carcinog

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“…The sampled cells gained from expertly selected tumor tissue, resemble the tumor with all its heterogeneity and follow-up data of the patient and a detailed pathology is available to allow the characteristics of the culture to be compared with those of the original tumor . This extensive genomic instability and variation in RNA expression during subculture was confirmed by Hiorns et al and provided supportive data for evidence that cell lines do evolve in culture, thereby weakening the direct relevance of such established cultures as models of human cancer 10 …”

Section: Introductionmentioning

confidence: 56%

Patient Derived Cell Culture and Isolation of CD133<sup>+</sup> Putative Cancer Stem Cells from Melanoma

Welte

Davies

Schäfer

et al. 2013

JoVE

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Despite improved treatments options for melanoma available today, patients with advanced malignant melanoma still have a poor prognosis for progression-free and overall survival. Therefore, translational research needs to provide further molecular evidence to improve targeted therapies for malignant melanomas. In the past, oncogenic mechanisms related to melanoma were extensively studied in established cell lines. On the way to more personalized treatment regimens based on individual genetic profiles, we propose to use patient-derived cell lines instead of generic cell lines. Together with high quality clinical data, especially on patient follow-up, these cells will be instrumental to better understand the molecular mechanisms behind melanoma progression.Here, we report the establishment of primary melanoma cultures from dissected fresh tumor tissue. This procedure includes mincing and dissociation of the tissue into single cells, removal of contaminations with erythrocytes and fibroblasts as well as primary culture and reliable verification of the cells' melanoma origin.Recent reports revealed that melanomas, like the majority of tumors, harbor a small subpopulation of cancer stem cells (CSCs), which seem to exclusively fuel tumor initiation and progression towards the metastatic state. One of the key markers for CSC identification and isolation in melanoma is CD133. To isolate CD133 + CSCs from primary melanoma cultures, we have modified and optimized the Magnetic-Activated Cell Sorting (MACS) procedure from Miltenyi resulting in high sorting purity and viability of CD133 + CSCs and CD133 -bulk, which can be cultivated and functionally analyzed thereafter.

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“…A variation of the gene expression during passaging was confirmed also in cell lines [43] and the authors warn that even comparisons of analyses of cell line cultures carrying the same name may be dangerous.…”

Section: Discussionmentioning

confidence: 94%

Effect of Different Factors on Proliferation of Antler Cells, Cultured In Vitro

et al. 2011

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Antlers as a potential model for bone growth and development have become an object of rising interest. To elucidate processes explaining how antler growth is regulated, in vitro cultures have been established. However, until now, there has been no standard method to cultivate antler cells and in vitro results are often opposite to those reported in vivo. In addition, many factors which are often not taken into account under in vitro conditions may play an important role in the development of antler cells. In this study we investigated the effects of the antler growth stage, the male individuality, passaged versus primary cultures and the effect of foetal calf serum concentrations on proliferative potential of mixed antler cell cultures in vitro, derived from regenerating antlers of red deer males (Cervus elaphus). The proliferation potential of antler cells was measured by incorporation of 3H thymidine. Our results demonstrate that there is no significant effect of the antler growth stage, whereas male individuality and all other examined factors significantly affected antler cell proliferation. Furthermore, our results suggest that primary cultures may better represent in vivo conditions and processes occurring in regenerating antlers. In conclusion, before all main factors affecting antler cell proliferation in vitro will be satisfactorily investigated, results of in vitro studies focused on hormonal regulation of antler growth should be taken with extreme caution.

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Variation in RNA expression and genomic DNA content acquired during cell culture

Cited by 30 publications

References 22 publications

Identification of possible genetic alterations in the breast cancer cell line MCF-7 using high-density SNP genotyping microarray

Identification of possible genetic alterations in the breast cancer cell line MCF-7 using high-density SNP genotyping microarray

Patient Derived Cell Culture and Isolation of CD133<sup>+</sup> Putative Cancer Stem Cells from Melanoma

Effect of Different Factors on Proliferation of Antler Cells, Cultured In Vitro

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