A TECHNIC FOR CHROMOSOME EVALUATION IN MAN, MONKEY, AND CELL CULTURE.
Progress rept. Dec 63-Aug 67,
SCHOOL OF AEROSPACE MEDICINE BROOKS AFB TEX
Pagination or Media Count:
When evaluating the chromosomes in man, plasma leukocytes are obtained by allowing the heparinized blood to stand at room temperature and those of the monkey are obtained from the buffy layer following centrifugation. Human leukocytes are seeded in TC medium 199 plus 0.25 ml. of Difco PHA M monkey leukocytes, in 70 NCTC 109, 20 fetal bovine serum, 10 F10 medium, and 0.1 ml. of pokeweed mitogen. Secondary cell cultures of human origin are not treated with mitogen because of their mitotic index. All cultures are incubated approximately 72 hours prior to gradient Colcemid treatments, followed by hypotonic treatments--distilled water for human leukocytes, 0.5 sodium citrate for monkey leukocytes and secondary mammalian cells. All cells are fixed with Carnoys fixative. Several drops of leukocyte suspension are placed on a chilled slide and ignited over a flame. The dried spreads are stained with either Giemsa or aceto-orcein. For mammalian cells, a few drops of aceto-orcein stain are mixed with the cell suspension and a drop of this suspension is placed on a microscopic slide and subjected to hand pressure under a coverslip. The coverslip is sealed with Sealit. Author