Chromosome Preparation From Cultured Cells

Bradley Howe; Ayesha Umrigar; Fern Tsien

doi:10.3791/50203

Chromosome Preparation From Cultured Cells

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Biology

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JoVE Journal Biology

Chromosome Preparation From Cultured Cells

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07:42 min

January 28, 2014

DOI: 10.3791/50203-v

Bradley Howe¹, Ayesha Umrigar¹, Fern Tsien¹

¹Department of Genetics,Louisiana State University Health Science Center

Overview

This article describes the procedure for harvesting and preparing chromosomes from live cells, including lymphocytes and skin fibroblasts. The prepared chromosomes can be utilized for various cytogenetic techniques such as G-banding and fluorescence in situ hybridization (FISH).

Key Study Components

Area of Science

Cytogenetics
Cell Biology
Molecular Biology

Background

Chromosomes can be isolated from live cells.
Common sources include human and mouse cells.
Chromosome preparations are essential for various cytogenetic analyses.
Techniques like G-banding and FISH are widely used in research.

Purpose of Study

To harvest chromosomes for G-banding and molecular cytogenetic tests.
To provide a reliable method for chromosome preparation.
To facilitate further analysis using advanced cytogenetic techniques.

Methods Used

Culture cells to logarithmic phase.
Harvest chromosomes using a hypotonic solution and fixative.
Drop cell suspension onto slides.
Stain slides for monitoring chromosomal preparation.

Main Results

Successful isolation of chromosomes from cultured cells.
Preparation suitable for G-banding and molecular techniques.
Effective monitoring of chromosomal integrity through staining.
Demonstrated applicability of the method for various cytogenetic analyses.

Conclusions

The described method provides a reliable approach for chromosome preparation.
It supports a range of cytogenetic techniques.
The procedure is applicable to both human and mouse cells.

Frequently Asked Questions

What types of cells can be used for chromosome isolation?

Chromosomes can be isolated from live cells such as lymphocytes and skin fibroblasts.

What techniques can be performed with the prepared chromosomes?

The prepared chromosomes can be used for G-banding, FISH, CGH, and SKY.

How are chromosomes harvested from cells?

Chromosomes are harvested using a hypotonic solution and a fixative after culturing the cells.

What is the purpose of staining the slides?

Staining helps monitor the chromosomal preparation and assess integrity.

Can this method be applied to both human and mouse cells?

Yes, the method is applicable to both human and mouse cells.

Chromosomes can be isolated from live cells such as lymphocytes or skin fibroblasts, and from organisms including humans or mice. These chromosome preparations can be further utilized for routine G-banding and molecular cytogenetic procedures such as fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH), and spectral karyotyping (SKY).

The overall goal of this procedure is to harvest and prepare chromosomes for GB banding and molecular cytogenetic tests. First, culture the cells to logarithmic phase. Then harvest the chromosomes with smid, a hypotonic and a fixative.

Drop the cell suspension into slides. Stain one slide. As a monitor of the chromosomal preparation, proceed to GB banding or molecular techniques of the remaining slides.

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