Premium
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
Author(s) -
Smith Stephen L.,
Kiss Joseph,
Siatskas Christopher,
Medin Jeffrey A.,
Moldwin Richard L.
Publication year - 2004
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2003.00661.x
Subject(s) - thrombopoietin , haematopoiesis , vascular endothelial growth factor , cord blood , cd34 , biology , growth factor , medicine , endocrinology , stem cell factor , microbiology and biotechnology , chemistry , immunology , stem cell , receptor , cancer research , biochemistry , vegf receptors
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.