Enhanced effect of vascular endothelial growth factor, thrombopoietin peptide agonist, SCF, and Flt3-L on LTC-IC and reporter gene transduction from umbilical cord blood CD34+ cells. Transfusion 2004 Mar;44(3):438-49
Date
03/05/2004Pubmed ID
14996204DOI
10.1111/j.1537-2995.2003.00661.xScopus ID
2-s2.0-1642313669 (requires institutional sign-in at Scopus site) 2 CitationsAbstract
BACKGROUND: Hemangioblastic precursors have been identified that give rise to both endothelial cells and HPCs, suggesting that common growth factor requirements may exist.
STUDY DESIGN AND METHODS: The effect of vascular endothelial growth factor (VEGF) in combination with thrombopoietin peptide agonist (TPOA), Flt-3 L (F), and SCF (S) on long-term culture-initiating cell (LTC-IC), CFU, differentiation, and transduction of cord blood (CB) CD34+ were evaluated up to 4 weeks in culture.
RESULTS: At Week 4, in cultures containing T/F/S and VEGF, the LTC-IC increased 1000-fold (from 37 +/- 8 to 37,012 +/- 14,329) with a frequency of 3.4 in 10,000 cells. In the T/F/S cultures without VEGF, the LTC-IC increased 675-fold (to 25,086 +/- 12,102) with a frequency of one LTC-IC in 10,000 cells. The addition of VEGF significantly increased (p < 0.05) the LTC-IC per 10,000 CB CD34+ cells. Transduction with reporter gene enhanced green fluorescent protein (EGFP), resulted in an increase in EGFP+ CFU at Week 1 and EGFP + LTC-IC at Weeks 1 and 4. The cells maintained their multilineage expression when cultured in conditions for erythroid, myeloid, or megakaryocytic differentiation. Peak percentage EGFP coexpression of GlyA and CD11b was 51 +/- 6 percent and 63 +/- 15 percent, respectively, at Week 2, while CD41a was 34 +/- 17 percent at Week 4.
CONCLUSION: T/F/S and VEGF have an enhanced effect on total LTC-IC output and frequency but do not appear to significantly alter the differentiation or transducibility characteristics of CB HPCs in vitro.
Author List
Smith SL, Kiss J, Siatskas C, Medin JA, Moldwin RLAuthor
Jeffrey A. Medin PhD Professor in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Antigens, CD34Cell Count
Cell Culture Techniques
Cell Differentiation
Cell Lineage
Cells, Cultured
Drug Synergism
Fetal Blood
Gene Expression
Green Fluorescent Proteins
Hematopoietic Stem Cells
Humans
Luminescent Proteins
Megakaryocytes
Membrane Proteins
Peptides
Retroviridae
Stem Cell Factor
Thrombopoietin
Transduction, Genetic
Vascular Endothelial Growth Factor A