Peer-Reviewed Journal Details
Mandatory Fields
Schmeckpeper, J;Ikeda, Y;Kumar, AHS;Metharom, P;Russell, SJ;Caplice, NM
2009
September
Differentiation
Lentiviral tracking of vascular differentiation in bone marrow progenitor cells
Validated
WOS: 16 ()
Optional Fields
SMOOTH-MUSCLE-CELLS HEMATOPOIETIC STEM-CELLS APOE-DEFICIENT MICE GROWTH-FACTOR VEGF TRANSGENIC MICE ENDOTHELIAL-CELLS BLOOD-VESSELS IN-VIVO FLUORESCENT PROTEIN TUMOR ANGIOGENESIS
78
169
176
Lentiviral vectors encoding for identifiable marker genes controlled by lineage-specific promoters can be used to track differentiation of bone marrow progenitors into endothelial cells and/or smooth muscle cells. Human VE-Cadherin and Smoothelin-B promoters were cloned into a self-inactivating lentiviral vector (HR-VECad and HR-SMTHB) and used to drive expression of green fluorescent protein (eGFP). These constructs demonstrated specific promoter activity in mature endothelial and smooth muscle cells respectively in vitro. Lin(-) bone marrow progenitor cells (Lin(-) BMCs) in culture were used to test vector ability to track vascular differentiation. HR-VECad transduced Lin(-) BMCs were plated on collagen-coated slides and grown in endothelial media, while HR-SMTHB transduced Lin(-) BMCs were cultured on fibronectin-coated slides and grown in smooth muscle media. For in vivo differentiation assessment, lentiviral transduced Lin(-) BMCs resuspended in Matrigel were injected subcutaneously into C57BL/6J mice. Explants were evaluated for eGFP expression. Lin(-) BMCs grown in endothelial differentiation media produced groups of polygonal endothelial-like cells by days 16-21. When transduced with HR-VEC advector, these expressed eGFP in distinct cells within the colony by days 18-21, and coexpressed VE-Cadherin and eNOS. Lin(-) BMCs grown in smooth muscle differentiation media produced spindle-shaped cells between days 10-14 in culture. When transduced with the HR-SMTHB vector, these cells showed eGFP expression at similar to 12 days, which increased over time and coexpressed alpha SMA, calponin and myosin heavy chain. Within Matrigel plugs containing HR-VECad transduced cells, eGFP(+) constituted 0.4 +/- 0.2% of total cells. In contrast, within Matrigel plugs containing HR-SMTHB transduced cells, eGFP(+) cells constituted 0.2 +/- 0.1% of total cells. These data demonstrate the feasibility of selectively marking BMC populations for cell fate determination. (C) 2009 International Society of Differentiation. Published by Elsevier Ltd. All rights reserved.
OXFORD
0301-4681
10.1016/j.diff.2009.01.002
Grant Details