Assessment of Modified Chromosome Analysis Method Using Culture Tube for Peripheral Blood

Lee, Young Kyung; Lee, Kwang Hee; Park, Ji Young; Cho, Hyoun Chan

Abstract

BACKGROUND: Peripheral blood is the most frequently used specimen for routine chromosome analysis. The classical method using flask for cell culture needs a lot of reagent and is cumbersome. To simplify the procedure, we tried using culture tube and modifying the fixation method. The purpose of this study is to assess the reliability of the modified method using culture tube for chromosome analysis of peripheral blood.
METHODS
We tested peripheral blood of ten normal healthy persons. In the modified method, culture tube (110 mm 16 mm, Nunc, Roskilde, Denmark) containing 2.25 mL RPMI 1640 supplemented with fetal calf serum and phytohemagglutinin was used. Harvest was done in the culture tube. Fixative 2.0 mL was added without centrifugation after hypotonic treatment. After 10 min incubation at room temperature, the cells were pelleted and washed. This method was compared with the reference method using flask. We evaluated the quality of chromosome and calculated mitotic index.
RESULTS
Chromosome quality of the modified method using culture tube was good and the same as that of the reference method. Mitotic index was higher in the modified method (1.0~4.3%, mean 2.5%) than in the reference method (0.4~2.2%, mean 1.2%) (P<0.01).
CONCLUSIONS
The modified method needs less amount of reagent and is easy to do. The chromosome quality was good enough to evaluate the karyotype. So, this modification enable to improve the effectiveness of chromosome laboratory.