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

Stephen L Smith, Joseph E Kiss, Christopher Siatskas, Jeff Medin, Richard L Moldwin

Research output: Contribution to journalArticleResearchpeer-review

2 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)438 - 449
Number of pages12
JournalTransfusion
Volume44
Issue number3
DOIs
Publication statusPublished - 2004
Externally publishedYes

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