Comparative studies of different stromal cell microenvironments in support of human B-cell development

• Published in: Experimental hematology 1999-08, Vol.27 (8), p.1271-1281
• Main Authors: Kurosaka, Daitaro, LeBien, Tucker W., Pribyl, Julie A.R.
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD19 - immunology; B cell; B-Lymphocytes - cytology; Bone Marrow Cells - drug effects; Bone Marrow Cells - physiology; Cell Adhesion Molecules - physiology; Cell Lineage; Cells, Cultured; Coculture Techniques; Culture Media, Serum-Free; DNA Nucleotidylexotransferase - biosynthesis; Fibroblasts - physiology; Flow Cytometry; Gene Expression; Gene Rearrangement, B-Lymphocyte; Hematopoiesis; Hematopoiesis - drug effects; Hematopoiesis - physiology; Hematopoietic Stem Cells - drug effects; Hematopoietic Stem Cells - physiology; Humans; Interleukin-3 - pharmacology; Mice; Reverse Transcriptase Polymerase Chain Reaction; Skin - cytology; Stem cell; Stromal cell; Stromal Cells - physiology
• ISSN: 0301-472X; 1873-2399
• DOI: 10.1016/S0301-472X(99)00067-3

This study compared human murine stromal cells for their capacity to support human hematopoietic stem cell (HSC) development into the B lineage. FACS sorted human fetal bone marrow (BM) HSC (CD34 +CD19 − or CD34 +/CD10 −/CD19 −/CD45RA −) were cultured on human fetal BM stromal cells, human skin fibroblasts, or murine S17 stromal cells and analyzed by flow cytometry or reverse transcriptase polymerase chain reaction. CD34 +CD19 − HSC on human BM stromal cells or fibroblasts differentiated into B-lineage cells with a continuum in density of surface CD19 expression, and some cells expressing μ/κ or μ/λ B-cell receptors. In contrast, CD19 + cells from S17 cultures had two- to fourfold higher levels of CD19, but no cells expressing B-cell receptors. The number and percentage of CD19 + cells was high, intermediate, or low in the human BM, human fibroblast, or murine S17 stromal cell cultures, respectively. Reverse transcriptase polymerase chain reaction analysis showed that TdT, CD19, and D HQ52-J H rearrangements were expressed at comparable levels when CD34 +/CD19 − HSC were plated on human or murine stromal cells. In contrast, CD34 +/CD10 −/CD19 −/CD45RA − HSC plated on human or murine stromal cells expressed CD19 in both cultures, but TdT was only expressed in human stromal cell cultures. We conclude that human BM stromal cell, human skin fibroblasts, and murine S17 stromal cell cultures can provide complementary and comparative tools for identification of stromal cell ligands with potentially unique functions in regulating human B-cell development.