Chromosome Preparation

Abstract

Standard chromosome preparations can be obtained from various tissues containing dividing cells. The easiest way to obtain metaphases for constitutional karyotyping is to take a blood sample and culture the lymphocytes. Amniotic fluid obtained by amniocentesis, or preparation of chromosomes from chorionic villi, may be used for prenatal diagnosis. Bone marrow is the tissue of choice for studies of acquired chromosome abnormalities in haematological malignancies. Chromosome preparations can also be made from cultures of solid tissues, like tumours, fetal skin, muscle, placenta, lung, kidney and liver. To have an unlimited source of cells, immortalised cell lines may be established, like tumour‐derived cell lines or cell transduction of normal cells with genes from a deoxyribonucleic acid (DNA) tumour virus. To achieve a higher resolution using fluorescent in situ hybridisation, methods for preparation of released, less‐condensed and linearised DNA fibres have been developed.

Key Concepts:

  • Metaphase spreads may be obtained from various tissues on condition that the cells are spontaneously dividing or may be cultured and in some cases chemically stimulated to divide by the addition of a mitogen.

  • The easiest way to obtain metaphase chromosomes for constitutional karyotyping is to take a blood sample and culture the lymphocytes.

  • Amniotic fluid or chorionic villi cells may be used for preparation of chromosomes for prenatal diagnosis.

  • Chromosome preparations from bone marrow may be used for studies of acquired chromosome abnormalities in haematological malignancies.

  • The cultures may be synchronised in order to achieve a better quality of chromosome preparation.

  • Chromosome preparations can be made from cultures of solid tissues after disaggregation.

  • To have an unlimited source of cells, immortalised cell lines may be established.

  • To achieve a higher resolution using fluorescent in situ hybridisation (FISH), methods for preparation of released, less‐condensed and linearised DNA fibres may be used.

Keywords: chromosome preparation; metaphase; cell culture; immortalised cell lines; fragile site; fibre‐FISH

Figure 1.

Principles of cell culture and standard chromosome preparation.

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References

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Further Reading

Heng HH, Windle B and Tsui L-C (2005) High‐resolution FISH analysis. Current Protocols in Human Genetics 44: 4.5.1–4.5.23.

Houseal TW, Dackowski WR, Landes GM and Klinger KW (1994) High resolution mapping of overlapping cosmids by fluorescence in situ hybridization. Cytometry 15: 193–198.

Senger G, Jones TA, Fidlerova H et al. (1994) Released chromatin: linearized DNA for high resolution fluorescence in situ hybridization. Human Molecular Genetics 3: 1275–1280.

Web Links

Fragile site, folic acid type, rare, fra(11)(q23.3) (FRA11B); Locus ID: 2439. LocusLink: http://www.ncbi.nlm.nih.gov/gene/?term=2439

Fragile site, folic acid type, rare, fra(11)(q23.3) (FRA11B); MIM number: 600651. OMIM:http://omim.org/entry/600651

Fragile site, folic acid type, rare, fra(X)(q27.3), (macroorchidism, mental retardation) (FRAXA); Locus ID: 2477. LocusLink: http://www.ncbi.nlm.nih.gov/gene/?term=2477

Fragile site, folic acid type, rare, fra(X)(q27.3), (macroorchidism, mental retardation) (FRAXA); MIM number: 309550. OMIM: http://omim.org/entry/309550

Fragile site, folic acid type, rare, fra(X)(q28) (FRAXE); Locus ID: 2481. LocusLink: http://www.ncbi.nlm.nih.gov/gene/?term=2481

Fragile site, folic acid type, rare, fra(X)(q28) (FRAXE); MIM number: 309548. OMIM: http://omim.org/entry/309548

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How to Cite close
Blennow, Elisabeth, and Iwarsson, Erik(Apr 2014) Chromosome Preparation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0005776.pub2]