Ex vivo expansion of CD34+ and T and NK cells from umbilical cord blood for leukemic BALB/C nude mouse transplantation

CBSCT with low incidence of GVHD is associated with higher rates of delayed engraftment and relapse compared with other stem cells transplants. The immune-mediated effect of NK and cytotoxic T cells against residual tumor cells was shown to prevent relapse and reinduce remission after bone marrow transplantation. We expanded CD34+ and T and NK cells ex vivo in cord blood with different cytokines combination and transplanted into leukemic BALB/C nude mouse. The results showed significant expansion of MNCs and CD34+ cells. The CD3+ T cells increased in the groups containing cytokines cocktail, especially in the group with IL-7 or IL-2. CD56+ NK cells number increased significantly only in a medium containing IL-2. Of the 20 engrafted BALB/C nude mice, 14 survived after 6 weeks transplantation, and the numbers in each group were from 3 to 4. Human CD3+ cells in the bone marrow of the survived mice were analyzed by flow cytometry and showed existing evidences. RT-PCR was used to detect leukemic fusion bcr/abl gene; all mice that experienced expanded cord blood transplantation could not be found to have expression of fusion bcr/abl gene. These suggest that T, NK cells as well as CD34+ cells could be expanded from CB MNCs in the same medium with the combination of cytokines. The expanded CB MNCs could reconstitute hematopoiesis and eliminate minimal residue leukemia disease in transplanted mice.