Megakaryocyte Differentiation and Platelet Formation from Human Cord Blood-derived CD34+ Cells

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2018 Jan 25

PMID 29364213

Citations 12

Authors

Jose Perdomo

Feng Yan

Halina H L Leung

Beng H Chong

Affiliations

Soon will be listed here.

Abstract

Platelet production occurs principally in the bone marrow in a process known as thrombopoiesis. During thrombopoiesis, hematopoietic progenitor cells differentiate to form platelet precursors called megakaryocytes, which terminally differentiate to release platelets from long cytoplasmic processes termed proplatelets. Megakaryocytes are rare cells confined to the bone marrow and are therefore difficult to harvest in sufficient numbers for laboratory use. Efficient production of human megakaryocytes can be achieved in vitro by culturing CD34 cells under suitable conditions. The protocol detailed here describes isolation of CD34 cells by magnetic cell sorting from umbilical cord blood samples. The necessary steps to produce highly pure, mature megakaryocytes under serum-free conditions are described. Details of phenotypic analysis of megakaryocyte differentiation and determination of proplatelet formation and platelet production are also provided. Effectors that influence megakaryocyte differentiation and/or proplatelet formation, such as anti-platelet antibodies or thrombopoietin mimetics, can be added to cultured cells to examine biological function.

Citing Articles

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