Reference List

Notes: (Language in Japanese)
Reference ID: 1215

Notes: Strain HeLa cells (Human cervical carcinoma) were established by Gey et al and have been distributed widely in the world. According to the selection of culture media and culture methods considerably wide range of substrains were proguced from the original line. Many works have been done on the properties of these cells until today. In the present paper the most popular type of HeLa cells distributed in Japan were examined of their fundamental conditions for cultivation in the simplified replicate tissue culture. And the substitution for serum proteins essential to the propagation was attempted with polyvinylpyrrolidone.
Reference ID: 2717

Notes: Genome Technology Branch, National Human Genome Research Institute, NIH, Bethesda, Maryland 20892, USA mmacville@patholaznnl
Abstract: We revisited the cytogenetic alterations of the cervical adenocarcinoma cell line HeLa through the use of spectral karyotyping (SKY), comparative genomic hybridization (CGH), and fluorescence in situ hybridization (FISH). SKY analysis unequivocally characterized all abnormal chromosomes. Chromosomal breakpoints were primarily assigned by simultaneous assessment of SKY painted chromosomes and inverted 4,6-diamidino2-phenylindole banding from the same cell. Twenty clonally abnormal chromosomes were found. Comparison with previously reported HeLa G-banding karyotypes revealed a remarkably stable cytogenetic constitution because 18 of 20 markers that were found were present before. The classification of 12 markers was refined in this study. Our assignment of the remaining six markers was consistent with those described in the literature. The CGH map of chromosomal copy number gains and losses strikingly matched the SKY results and was, in a few instances, decisive for assigning breakpoints. The combined use of molecular cytogenetic methods SKY, CGH, and FISH with site-specific probes, in addition to inverted 4,6-diamidino-2-phenylindole or conventional G-banding analysis, provides the means to fully assess the genomic abnormalities in cancer cells. Human papillomaviruses (HPVs) are frequently integrated into the cellular DNA in cervical cancers. We mapped by FISH five HPV18 integration sites: three on normal chromosomes 8 at 8q24 and two on derivative chromosomes, der(5)t(5;22;8)(qll;q11q13;q24) and der(22)t(8; 22)(q24;q13), which have chromosome 8q24 material. An 8q24 copy number increase was detected by CGH. Dual-color FISH with a c-MYC probe mapping to 8q24 revealed colocalization with HPV18 at all integration sites, indicating that dispersion and amplification of the c-MYC gene sequences occurred after and was most likely triggered by the viral insertion at a single integration site. Numerical and structural chromosomal aberrations identified by SKY, genomic imbalances detected by CGH, as well as FISH localization of HPV18 integration at the c-MYC locus in HeLa cells are common and representative for advanced stage cervical cell carcinomas. The HeLa genome has been remarkably stable after years of continuous cultivation; therefore, the genetic alterations detected may have been present in the primary tumor and reflect events that are relevant to the development of cervical cancer
Reference ID: 5247

Notes: Culture media for mammalian cells contain electrolytes at concentrations that approximate those of the extracellular fluid (ECF). However, we observed that HeLa strain cells can grow in a cytosol type, serum-free, high amino acid medium (KN13) in which the concentrations of K+, Mg2+ and Pi are essential to preent the harmful effect of a high concentration of K+ in the medium. Even an increase in the K/Na ratio to 7 and 9 permitted growth for at least 2 weeks and 4 days, respectively. HeLa P3 cells adapted to KN13 (HeLa K cells) have proliferated (4 fold/week) for 4 years in this medium. The Na, K-ATPase of HeLa K cells was the alpha1 isoform. HeLa K cells in KN13 meium have alower membrane potential (-3.0 +- 6.1mV) than that of the original HeLa P3 cells in DM160 (-26 +- 10mV).
Reference ID: 5328