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.
