Metaphase synchronization and chromosome preparation from the OK opossum cell line having a potentially isolatable X chromosome

Abstract

As part of a study on X chromosomes, metaphase cell synchrony and chromosome isolation methods were developed for the opossum (Didelphis virginiana) kidney epithelial cell line (OK). The cell synchrony yielded large amounts of metaphase cells using a relatively simple method in which a key feature was a calcium- and magnesium-free balanced salt wash. A neutral pH chromosome isolation method was developed for the kidney epithelial cells, because they were somewhat difficult to disrupt fully by other methods. FACS IV flow microfluorometric analysis of OK chromosomes confirms a clear difference between the sizes of opossum X chromosomes and autosomes.

Growth Inhibition and Chromosomal Instability of Cultured Marsupial (Opossum) Cells after Treatment with DNA Polymerase α Inhibitor

“…OK cells established from Didelphis marsupialis virginiana show moderate mixoploidy, with 22, 23, or 24 chromosomes. 20) Almost all of the OK cells used in this study had 24 chromosomes (Fig. 3A).…”