Background: Hematopoiesis, is all the cells within the blood system being formed in a specific hierarchy from a small community of hematopoietic stem cells. The aim of this study was to identify the Tbx3 protein levels at stages of physiological development of blood cells in hematopoiesis. Methods: The isolation of hematopoietic stem cell (HSC) was performed from the umbilical cord blood and cultured for differentiation to myeloid series for 14 days. The expression levels of CD34, CD13, CD45, ALDH (aldehyde dehydrogenase) in hematopoietic progenitor cells isolated from cord blood and myeloid series were checked. The levels of Tbx3 in myeloid series and HSC analyzed with flow cytometry. For statistical analysis student’s t test was used and p values ≤ 0.05 were considered statistically significant. Results: CD34, CD45 and ALDH levels were high in hematopoietic progenitor cells. After the differentiation CD13 levels increased, whereas CD34, CD45, ALDH levels were decreased. Tbx3 protein was shown in the hematopoietic stem cells obtained from umbilical cord blood and myeloid series cells. There was no significant difference between Tbx3 protein levels of the umbilical cord blood and myeloid series cells. Conclusion: The presence of Tbx3 transcription factor in hematopoiesis has been shown for the first time with this study.