TY  - JOUR
TI  - Isolation of Human Umbilical Cord Blood Hematopoietic Stem Cells and Directed Differentiation Into Megakaryocytes
AU  - Xia, Wenjun 
AU  - Miao, Zeqing 
AU  - Zhang, Weiwei 
AU  - Liu, Zhixia 
AU  - Wu, Huang 
AU  - Fan, Yahan 
VL  - 16
IS  - 19
PY  - 2026
DA  - 2026/10/05
SP  - e5822
C1  - Bio-protocol 2026;16:e5822
DO  - 10.21769/BioProtoc.5822
UR  - https://doi.org/10.21769/BioProtoc.5822
AB  - Platelets originate from megakaryocytes, whose generation involves a series of biological processes including directed differentiation, proliferation, polyploidization, and maturation of hematopoietic stem cells. Abnormalities in megakaryocyte development and maturation can lead to quantitative and functional defects in platelets, thereby contributing to hemostatic or thrombotic disorders as well as the development of malignancies. Investigating megakaryocyte development and maturation and platelet production can provide important theoretical foundations for the diagnosis and treatment of thrombocytopenia, thrombotic diseases, and myeloproliferative neoplasms. Currently, there are three main clinical sources of hematopoietic stem cells (HSCs): bone marrow (BM), peripheral blood (PBSC), and umbilical cord blood (UCB). Among these, umbilical cord blood (UCB)-derived HSCs, due to their higher differentiation efficiency and stronger proliferative capacity, are the preferred starting cell source for studying megakaryocyte (MK) development and maturation and the mechanisms of platelet production. This article describes a detailed protocol covering all necessary steps for isolating CD34+ hematopoietic stem cells from umbilical cord blood, followed by in vitro induction culture with stem cell factor (SCF) and thrombopoietin (TPO) to generate mature megakaryocytes that highly express early megakaryocyte markers (CD41a, CD61) and late maturation markers (CD42a, CD42b). This protocol provides an effective tool for studying megakaryocyte development and platelet production and holds potential value for application in research on megakaryocyte-related diseases.
KW  - Umbilical cord blood
KW  - Hematopoietic stem cells
KW  - Megakaryocyte differentiation
KW  - Thrombopoiesis
KW  - CD34+
KW  - SCF
KW  - TPO
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False