Preparation and culture of products of conception and other solid tissues for chromosome analysis.
In clinical settings, chromosome studies are usually performed on solid tissue other than solid tumors for one of two reasons: the tissue biopsy is the only tissue available from the patient, or tissues other than the standard peripheral blood lymphocytes must be examined because of suspected mosaicism. This unit describes methods for culturing tissue samples to be used for preparation of metaphase chromosomes or for biochemical or DNA analysis. Protocols include an efficient enzymatic cell disruption procedure and a mechanical disruption procedure. In addition, a procedure for processing uncultured cells for analysis by fluorescence in situ hybridization (FISH) is provided. Finally, a method is presented that can sometimes be applied to placental tissue involving the use of mitotically active chorionic villus tissue to prepare metaphase chromosome spreads directly, without cell culture. Two support protocols describe methods for collecting tissue samples.
Levy
Brynn
B
College of Physicians and Surgeons of Columbia University, New York, USA.
Jobanputra
Vaidehi
V
Warburton
Dorothy
D
D001706:Biopsy / Q000379:methods*; D020732:Cytogenetic Analysis / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D010920:Placenta / Q000166:cytology*; D046509:Tissue Culture Techniques / Q000379:methods*