Gene expression of human hematopoietic stem cells (CD34+) stimulated with native & mutant (244) form of myeloid stem-cell transcription factor PU.1 alone & in combination with myeloid specific cytokines (IL3+GMCSF+Flt2+MCSF+SCF)

Human CD34+ hematopoietic stem cells gives rise to complete blood lineages through the process of hematopoiesis. Here we tried to differentiate a particular lineage i.e myeloid from CD34+ hematopoietic stem cells using a transcription factor PU.1 and its mutated form. Gene expression analysis was performed after treating cells with protein factors and different cytokine combinations. Cells were given six different treatments including control-PU.1 only and PU.1 with cytokines combination; Y244D only and Y244D with cytokines combination and only cytokine combination (IL3, GMCSF, Flt2, MCSF, and SCF). After 5 days of incubation, cells were washed and pellet was preserved in RNAlater (Ambion) to be shipped to Genotypic Technologies Pvt. Ltd. (Bangalore) for microarray analysis. HSC Cells were given six different treatments including control-PU.1 only and PU.1 with cytokines combination; Y244D only and Y244D with cytokines combination and only cytokine combination (IL3, GMCSF, Flt2, MCSF, and SCF). After 5 days of incubation, cells were washed and pellet was preserved in RNAlater (Ambion) to be shipped to Genotypic Technologies Pvt. Ltd. (Bangalore) for microarray analysis.