A rapid banding technique for human chromosomes
About: This article is published in The Lancet.The article was published on 1971-10-30. It has received 4888 citations till now. The article focuses on the topics: G banding & Karyotype.
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TL;DR: Rapidly accumulating evidence indicates that structural variants can comprise millions of nucleotides of heterogeneity within every genome, and are likely to make an important contribution to human diversity and disease susceptibility.
Abstract: The first wave of information from the analysis of the human genome revealed SNPs to be the main source of genetic and phenotypic human variation. However, the advent of genome-scanning technologies has now uncovered an unexpectedly large extent of what we term 'structural variation' in the human genome. This comprises microscopic and, more commonly, submicroscopic variants, which include deletions, duplications and large-scale copy-number variants - collectively termed copy-number variants or copy-number polymorphisms - as well as insertions, inversions and translocations. Rapidly accumulating evidence indicates that structural variants can comprise millions of nucleotides of heterogeneity within every genome, and are likely to make an important contribution to human diversity and disease susceptibility.
1,804 citations
TL;DR: Three patients with hematologic relapse after bone marrow transplantation for chronic myelogenous leukemia were treated with interferon alpha and transfusion of viable donor buffy coat and had complete hematological and cytogenetic remission, which persisted 32 to 91 weeks after treatment, an example of adoptive immunotherapy without cytoreductive chemotherapy or radiotherapy in human chimeras.
1,419 citations
Additional excerpts
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Journal Article•
TL;DR: Two sublines of a breast epithelial cell culture, MCF-10, derived from human fibrocystic mammary tissue exhibit immortality after extended cultivation in low calcium concentrations, although later passages showed minimal rearrangement and near-diploidy, the immortal cells were not karyotypically normal.
Abstract: Two sublines of a breast epithelial cell culture, MCF-10, derived from human fibrocystic mammary tissue exhibit immortality after extended cultivation in low calcium concentrations (0.03-0.06 mM) and floating transfers in low calcium (MCF-10F), or by trypsin-Versene passages in the customary (normal) calcium levels, 1.05 mM (MCF-10A). Both sublines have been maintained as separate entities after 2.3 years (849 days) in vitro and at present have been in culture for longer than 4 years. MCF-10 has the characteristics of normal breast epithelium by the following criteria: (a) lack of tumorigenicity in nude mice; (b) three-dimensional growth in collagen; (c) growth in culture that is controlled by hormones and growth factors; (d) lack of anchorage-independent growth; and (e) dome formation in confluent cultures. Cytogenetic analysis prior to immortalization showed normal diploid cells; although later passages showed minimal rearrangement and near-diploidy, the immortal cells were not karyotypically normal. The emergence of an immortal culture in normal calcium media was not an inherent characteristic of the original tissue from which MCF-10 was derived since reactivated cryo-preserved cells from cultures grown for 0.3 and 1.2 years in low calcium were incapable of sustained growth in normal calcium.
1,410 citations
TL;DR: It is shown here that expression of a mutant catalytic subunit of human telomerase results in complete inhibition of telomer enzyme activity, reduction in telomere length and death of tumor cells, validating human telomersase reverse transcriptase as an important target for the development of anti-neoplastic therapies.
Abstract: Telomerase is a ribonucleoprotein enzyme that maintains the protective structures at the ends of eukaryotic chromosomes, called telomeres. In most human somatic cells, telomerase expression is repressed, and telomeres shorten progressively with each cell division. In contrast, most human tumors express telomerase, resulting in stabilized telomere length. These observations indicate that telomere maintenance is essential to the proliferation of tumor cells. We show here that expression of a mutant catalytic subunit of human telomerase results in complete inhibition of telomerase activity, reduction in telomere length and death of tumor cells. Moreover, expression of this mutant telomerase eliminated tumorigenicity in vivo. These observations demonstrate that disruption of telomere maintenance limits cellular lifespan in human cancer cells, thus validating human telomerase reverse transcriptase as an important target for the development of anti-neoplastic therapies.
1,048 citations
TL;DR: TF‐1 is a cell line of immature erythroid origin that requires GM‐CSF, IL‐3, or EPO for its growth and that has the ability to undergo differentiation into either more mature erystroid cells or into macrophage‐like cells.
Abstract: We have established a novel cell line, designated as TF-1, from a patient with erythroleukemia, which showed complete growth dependency on granulocyte-macrophage colony-stimulating factor (GM-CSF) or on interleukin-3 (IL-3) and carried a homogeneous chromosomal abnormality (54X). Erythropoietin (EPO) also sustained the short-term growth of TF-1, but did not induce erythroid differentiation. These three hematopoietic growth factors acted on TF-1 synergistically. Transforming growth factorβ and interferons inhibited the factor-dependent growth of TF-1 cells in a dose-dependent fashion, and monocyte-colony stimulating factor and interkeukin-1 enhanced the GM-CSF-dependent growth of TF-1. Ultrastructural studies revealed some very immature features in this cell line. Although TF-1 cells do not express glycophorin A or carbonyl anhydrase I, the morphological and cytochemical features, and the constitutive expression of globin genes, indicate the commitment of TF-1 to erythroid lineage. When induced to differentiate, TF-1 entered two different pathways. Specifically, hemin and delta-arninolevulinic acid induced hemoglobin synthesis, whereas TPA induced dramatic differentiation of TF-1 into macrophage-like cells. In summary, TF-1 is a cell lineof immature erythroid origin that requires GM-CSF, IL-3, or EPO for its growth and that has the ability to undergo differentiation into either more mature erythroid cells or into macrophage-like cells. TF-1 is auseful tool for analyzing the human receptors for IL-3, GM-CSF, and EPO or the signal transduction of these hemopoietic growth factors.
833 citations
References
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TL;DR: It seems probable, therefore, that the darker staining with Giemsa of these regions, after denaturation and annealing, indicates the presence of highly repetitive DNA.
Abstract: A GIEMSA staining procedure that preferentially stains centromeric heterochromatin in mouse chromosomes has been described1. This specificity was observed when fixed preparations were treated with sodium hydroxide to denature the DNA, and then incubated in warm saline to allow annealing, in the presence of 3H-labelled single stranded satellite DNA or its complementary RNA. In this way mouse satellite DNA was located in the centromeric heterochromatin1,2. It is known to consist of highly repetitive sequences3 and to anneal much more rapidly than non-repetitive DNA4. It seems probable, therefore, that the darker staining with Giemsa of these regions, after denaturation and annealing, indicates the presence of highly repetitive DNA.
1,140 citations
TL;DR: A convenient measuring technique has been developed for the rapid and accurate recording of fluorescence patterns in human metaphase chromosomes that should be particularly valuable for the identification of chromosomes 4–5 and the individual types in the 6–12 group.
Abstract: The distribution of DNA along metaphase chromosomes that are not excessively contracted can be visualized in the fluorescence microscope with the aid of fluorescent DNA-binding agents. Additional, characteristic details in the fluorescence patterns are obtained with fluorochromes that bind preferentially to certain chromosomal regions. The highly fluorescent alkylating agent quinacrine mustard (QM) effects discrete, fluorescent labeling of both plant and mammalian metaphase chromosomes, presumably by selective binding to guanine residues in DNA, and is also capable of intercalation in the DNA double helix. Chromosome regions fluorescing particularly strongly with QM have been demonstrated in human metaphase chromosomes 3, 13–15 and Y. A convenient measuring technique has been developed for the rapid and accurate recording of fluorescence patterns in human metaphase chromosomes. These photoelectric recordings of the fluorescence patterns contain far greater detail than can be seen by the human eye. The fluorescence patterns described are based on measurements of about 1,000 human metaphase chromosomes. This new technique of determining fluorescence patterns in human chromosomes should be particularly valuable for the identification of chromosomes 4–5 and the individual types in the 6–12 group. Individual, typical patterns also occur within the groups 13–15, 17–18, and 21–22.
919 citations
TL;DR: This method, which is based on treatment of the chromosomes in situ with NaOH, followed by incubation in sodium chloride-trisodium citrate and Giemsa staining, results in highly specific banding patterns in characteristic regions of the chromosome arms.
Abstract: Individual pairs of human chromosomes can be reliably identified by a new method that does not require special optical equipment and that results in permanent preparations. This method, which is based on treatment of the chromosomes in situ with NaOH, followed by incubation in sodium chloride-trisodium citrate and Giemsa staining, results in highly specific banding patterns in characteristic regions of the chromosome arms. It should prove useful for the detection of small structural changes in chromosomes.
262 citations
TL;DR: Using the Giemsa stain, it was observed that these areas stained more darkly than the remainder of the chromosome material on denaturation and reassociation in situ, indicating that, even in these conditions, there is re-association of the satellite DNA strands and that there may be a simpler technique in which no satellite DNA need be added.
Abstract: A SIMPLE technique is described here which locates regions of satellite or highly repetitive DNA within mammalian metaphase chromosomes and helps the study of relationships between these regions and those which are heterochromatic and late replicating. The technique arose from attempts to hybridize radioactively labelled satellite DNA from mouse1, guineapig2 and calf3 in situ with complementary regions in metaphase chromosomes, as described by Jones4 and Pardue and Gall5. The areas of hybrids formed in this manner were always located around the centromere. Using the Giemsa stain we also observed that these areas stained more darkly than the remainder of the chromosome material on denaturation and reassociation in situ, without the addition of satellite DNA. This indicated that, even in these conditions, there is re-association of the satellite DNA strands and that there may be a simpler technique in which no satellite DNA need be added. Ideally, a clear picture of the satellite DNA would be obtained in metaphase chromosomes by denaturing the DNA strands and allowing them to reassociate for varying times in an adequate buffer. In this manner, with increasing time of reassociation there would be within the chromosomes a progression of darkly staining regions of satellite DNA which would contrast sharply with lightly staining regions of the remaining DNA.
159 citations
TL;DR: A method has been developed which can provide further information about chromosome organization and is believed to locate repetitive DNA sequences in cytological chromosome preparations.
Abstract: STUDIES of mammalian chromosomes have advanced rapidly since the introduction of the quinacrine fluorescence technique1 and other procedures which are believed to locate repetitive DNA sequences in cytological chromosome preparations2,3. To improve these techniques, a method has been developed which can provide further information about chromosome organization.
116 citations