Monopolar preparation of human lymphocytes for evaluation of the metaphase chromosome alignment

概要

論文の詳細を見る
The metaphase alignment of chromosomes to the midzone is indispensable for the faithful segregation of sister chromatids in the process of cell division. In the present study, we describe a new method for the collection of monopolar-arrested human lymphocytes by a combination of monastrol treatment and low speed centrifugation (spin-down). In this preparation, cells with non-aligned chromosomes are easily discernible from those with complete alignment of chromosomes. Immunostaining of spindle and kinetochores is helpful for the accurate judgment whether chromosome alignment is complete or not. In non-immortalized lymphocytes (T-cells) and an immortalized lymphoblastoid (B-cells) cell line derived from control adults, greater than approximately 80% of cells among the metaphase population showed complete alignment of chromosomes by the monopolar/spin-down preparation. In contrast, incomplete alignment was found in 35-55% cells in lymphocytes derived from a Cornelia de Lange syndrome patient with a mutation in NIPBL gene. Homologues of NIPBL in yeast (Scc2) and Drosophia (Nipped-B) are required for sister chromatid cohesion during mitosis as a regulator of the binding of recently accentuated by the detection of congenital disorders with mutations in genes involved in chromosome assembly and segregation, the monopolar/spin-down preparations using human lymphocytes may be a useful tool for the evaluation of the quality of chromosome dynamics in the clinical context of these disorders.
財団法人染色体学会の論文

Monopolar preparation of human lymphocytes for evaluation of the metaphase chromosome alignment

スポンサーリンク

概要

著者

関連論文

スポンサーリンク