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Splenic Endothelial Cell Lines Support Development of Dendritic Cells from Bone Marrow

School of Biochemistry and Molecular Biology, The Australian National University, Canberra, Australian Capital Territory, Australia

Key Words. Spleen • Endothelial cells • Dendritic cell development • Microenvironment

Correspondence: Professor Helen C. O’Neill, B.Sc., Ph.D. School of Biochemistry and Molecular Biology, Building 41, Linnaeus Way, Australian National University, Canberra, ACT, Australia, 0200. Telephone: +61 2 6125 4720; Fax: +61 2 6125 0313; e-mail: helen.oneill{at}anu.edu.au

Received August 26, 2005; accepted for publication January 26, 2006.

Although growth factors are commonly used to generate dendritic cells (DCs) in vitro, the role of the microenvironment necessary for DC development is still poorly understood. The mixed splenic stromal cell population STX3 defines an in vitro microenvironment supportive of DC development. Dissection of cellular components of the STX3 stroma should provide information about a niche for DC development. STX3 was therefore cloned by single-cell sorting, and a panel of 102 splenic stromal cell lines was established. Four representative splenic stromal cell lines that support hematopoiesis from bone marrow are described here in terms of stromal cell type and DC production. All four stromal lines express the endothelial genes Acvrl1, Cd34, Col18a1, Eng, Flt1, Mcam, and Vcam1 but not Cd31 or Vwf. Three of the four lines form tube-like structures when cultured on Matrigel. Their endothelial maturity correlates with the ability to support myeloid DC development from bone marrow. A fourth cell line, unable to form tube-like structures in Matrigel, produced large granulocytic cells expressing CD11b and CD86 but not CD11c and CD80. Conditioned media from splenic stromal cell lines also support DC production, indicating that soluble growth factors and cytokines produced by stromal lines drive DC development. This article reports characterization of immature endothelial cell lines derived from spleen that are supportive of DC development and predicts the existence of such a cell type in vivo which regulates DC development within spleen.

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