1. Human brain endothelial cells (HUBEC) promote SCID repopulating cell expansion through direct contact
- Author
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Barry Meisenberg, Douglas K. Tadaki, Xiangfei Cheng, Emily Welty, Naoko Takebe, Ann M. Farese, and Thomas J. MacVittie
- Subjects
Clinical Biochemistry ,CD34 ,Antigens, CD34 ,Mice, SCID ,Biology ,CD38 ,Wnt3 Protein ,Mice ,Endocrinology ,Cell Adhesion ,medicine ,Animals ,Humans ,Receptor ,Cells, Cultured ,beta Catenin ,Cell Proliferation ,WNT Family Protein ,Wnt signaling pathway ,Brain ,Endothelial Cells ,Hematopoietic stem cell ,Cell Biology ,Hematopoietic Stem Cells ,ADP-ribosyl Cyclase 1 ,Coculture Techniques ,Cell biology ,Wnt Proteins ,medicine.anatomical_structure ,Immunology ,Intercellular Signaling Peptides and Proteins ,SCID-Repopulating Cell ,Severe Combined Immunodeficiency ,Endothelium, Vascular ,Proto-oncogene tyrosine-protein kinase Src - Abstract
The objective of this study was to re-evaluate the previously published hematopoietic stem cell (HSC) expansion work using human brain endothelial cells (HUBEC). The expansion effect of contact and non-contact conditions was reported to be equivalent by others. However, we report here different results that the expansion can be achieved only with direct contact. We co-cultured human CD34+ cells with and without HUBEC contact for seven days with cytokines and the readouts were CD34+ / CD38 - phenotype and SCID repopulating cell (SRC) frequency. Also tested was the inhibitory effect of Wnt receptor inhibitor Dkk-1 on HUBEC contact ex vivo expansion; whether an increased expression of Wnt3 occurs on the HUBEC surface; and detection of an increased nuclear localization of beta-catenin in CD34+ / CD38- cells in HUBEC contact culture condition. We conclude that the successful expansion by HUBEC contact culture is a candidate explanation based on the Wnt family protein, possibly Wnt3, expression on HUBEC.
- Published
- 2007
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