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Vol. 11, Issue 12, 4295-4308, December 2000
-binding Protein-1
Promotes In Vitro Differentiation of Embryonic Stem Cells into
Endothelium
¶¶
§¶
Departments of *Cell Biology, The latent transforming growth factor-Medicine, and ¶Kaplan Cancer
Center, New York University School of Medicine, New York, New
York 10016-6497
-binding protein-1
(LTBP-1) belongs to a family of extracellular glycoproteins that includes three additional isoforms (LTBP-2, -3, and -4) and the matrix
proteins fibrillin-1 and -2. Originally described as a TGF--masking
protein, LTBP-1 is involved both in the sequestration of latent TGF-
in the extracellular matrix and the regulation of its activation in the
extracellular environment. Whereas the expression of LTBP-1 has been
analyzed in normal and malignant cells and rodent and human tissues,
little is known about LTBP-1 in embryonic development. To address this
question, we used murine embryonic stem (ES) cells to analyze the
appearance and role of LTBP-1 during ES cell differentiation. In vitro,
ES cells aggregate to form embryoid bodies (EBs), which differentiate
into multiple cell lineages. We analyzed LTBP-1 gene expression and
LTBP-1 fiber appearance with respect to the emergence and distribution
of cell types in differentiating EBs. LTBP-1 expression increased
during the first 12 d in culture, appeared to remain constant
between d 12 and 24, and declined thereafter. By immunostaining,
fibrillar LTBP-1 was observed in those regions of the culture
containing endothelial, smooth muscle, and epithelial cells. We found
that inclusion of a polyclonal antibody to LTBP-1 during EB
differentiation suppressed the expression of the endothelial specific
genes ICAM-2 and von Willebrand factor and delayed the organization of
differentiated endothelial cells into cord-like structures within the
growing EBs. The same effect was observed when cultures were treated
with either antibodies to TGF- or the latency associated peptide, which neutralize TGF-. Conversely, the organization of endothelial cells was enhanced by incubation with TGF-1. These results suggest that during differentiation of ES cells LTBP-1 facilitates endothelial cell organization via a TGF--dependent mechanism.
Present addresses: Institute for Cancer
Research and Treatment, Department of Genetics, 10060 Candiolo
(Torino), Italy .
§
Corresponding author. E-mail address:
rifkid01{at}med.nyu.edu.
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