24-Hour Lymphocyte Culture Method: A Novel Way to Yield Metaphase Chromosomes

Abstract

The study aims to find a shorter incubation time and an alternative culture medium than the standard 96 hour lymphocyte culture method. Human umbilical cord blood was cultured for one, two, and three hours using RPMI 1640 with PHA, and for one hour using Ham’s F10 Nutrient Mixture and MarrowMAX. However, all cultures failed to yield metaphase chromosomes. The incubation period was then set to 24 hours using three media: Ham’s F10 Nutrient Mixture, RPMI 1640 with PHA, and RPMI 1640 without PHA. Cultures incubated overnight yielded metaphase chromosomes. Cultures in RPMI 1640 medium without PHA yielded the most number of metaphase spreads, followed by RPMI 1640 with PHA, and then by Ham’s F10 Nutrient Mixture. The method can be used for preliminary cytogenetic analysis, provided that a backup is prepared. This leads to better treatment strategies for patients with suspected cytogenetic disorders.