Korean J Hematol.  2003 May;38(2):81-90.

Differentiation and Apoptosis of the Erythroid, Myeloid and Megakaryocytic Lineage Cells in vitro Culture of Cord Blood CD34+ Cells Using Various Cytokines

• Ryu KH
• Park JY
• Cho SJ
• Seoh JY
• Kie JH

Abstract

BACKGROUND
In vitro generated cells from cord blood (CB) CD34+ cells increase cell dose and may reduce the severity and the duration of pancytopenia after transplantation. But safe engraftment for adolescents and adults is still not predictable and standardized. We attempted to establish a clinically application of in vitro generated cells by investigating the use of cytokines for the culture of CB CD34+ cells.
METHODS
CD34+ cells, purified from four separate human CB units by magnetic bead selection, were cultured in IMDM with several cytokines. Differentiation of the in vitro generated cells has been confirmed by flowcytometry with specific erythroid, myeloid and megakaryocytic lineage surface markers. And apoptosis of these cells also was analysed with annexin V staining and morphologic analysis under the electron microscopic examination was done, simultaneously.
RESULTS
Purified CB CD34+ cells were expanded with differentiation from CD38- to CD38+ expression with time. CB CD34+ cells could be terminally differentiated into erythroid (GPA+) and myeloid (CD33+/CD15+) lineage cells without apoptosis (annexin-V - ) in the presence of EPO and G-CSF, respectively. Megakaryocytic differentiation was partially arrested in early stage due to apoptosis in the presence of TPO. Morphological examination using electron microscope revealed all stages of erythroid and myeloid lineage cells without apoptosis, and apoptotic megakaryocytic lineage cells of early stage. But we could observe the small number of fully maturated platelets.
CONCLUSION
CB CD34+ cells were terminally differentiated to erythroid, myeloid, and megakaryocytic lineage cells with or without apoptosis by several cytokines.

Keyword

In vitro generated cells; Differentiation; Apoptosis; Cord blood CD34+ cells

MeSH Terms

Adolescent
Adult
Annexin A5
Apoptosis*
Cytokines*
Fetal Blood*
Granulocyte Colony-Stimulating Factor
Humans
Pancytopenia
Annexin A5
Cytokines
Granulocyte Colony-Stimulating Factor