Faculty Collaboration Database

Expansion of megakaryocyte precursors and stem cells from umbilical cord blood CD34+ cells in collagen and liquid culture media. J Hematother Stem Cell Res 2001 Jun;10(3):391-403

Date

07/17/2001

Pubmed ID

11454314

DOI

10.1089/152581601750288993

Scopus ID

2-s2.0-0034912524 (requires institutional sign-in at Scopus site)   11 Citations

Abstract

Umbilical cord blood (UCB) is now commonly used as a source of stem cells for hematopoietic reconstitution following myeloablative therapy in patients with a variety of diseases. Although UCB is a rich source of stem cells, platelet engraftment occurs at a median of 71 days which is significantly prolonged compared to allogeneic bone marrow. The number of megakaryocyte (MK) precursors in stem cell harvests appears to correlate inversely with the time to platelet engraftment. In an effort to increase the number of platelet precursors, we cultured CD34-selected cord blood mononuclear cells (MNC) in serum-free collagen medium with numerous cytokine combinations. The cells were cultured with four cytokines: interleukin-3 (IL-3), thrombopoietin (TPO), stem cell factor (SCF), and Flt-3); five cytokines, IL-3, TPO, SCF, Flt-3 plus granulocyte-macrophage colony-stimulating factor (GM-CSF), or erythropoietin (Epo); or all six cytokines in combination. After 16 days, significant expansion of MK precursors (CD41(+)) and stem cells (CD34(+) and AC133(+) cells) were seen in cells cultured in IL-3, TPO, SCF, and Flt-3 with or without GM-CSF compared to the combinations that contained Epo (p < 0.05). Similar studies were performed using liquid culture medium, and after 14 days the number of MNCs, CD34(+), AC133(+), CD41(+), and CD61(+) cells were higher in the UCB cells cultured in IL-3, TPO, SCF, and Flt-3 compared to those cultured with those four cytokines plus GM-CSF. These results demonstrate that UCB stem cells can be effectively expanded ex vivo and enriched with platelet precursors using TPO, SCF, Flt-3, and IL-3, whereas the addition of Epo and GM-CSF is unnecessary.

Author List

Shaw PH, Olszewski M, Kletzel M

Author

Peter H. Shaw MD Professor in the Pediatrics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

AC133 Antigen
Antigens, CD
Antigens, CD34
Cell Culture Techniques
Cell Division
Cells, Cultured
Collagen
Culture Media
Culture Media, Serum-Free
Cytokines
Drug Synergism
Erythropoietin
Fetal Blood
Glycoproteins
Granulocyte-Macrophage Colony-Stimulating Factor
Hematopoietic Cell Growth Factors
Hematopoietic Stem Cells
Humans
Immunomagnetic Separation
Integrin beta3
Interleukin-3
Megakaryocytes
Peptides
Platelet Glycoprotein GPIIb-IIIa Complex
Platelet Membrane Glycoproteins
Proto-Oncogene Proteins
Receptor Protein-Tyrosine Kinases
Stem Cell Factor
Thrombopoietin
fms-Like Tyrosine Kinase 3