Kurosaka D, LeBien T W, Pribyl J A
Department of Internal Medicine, The Jikei University of Medicine, Tokyo, Japan.
Exp Hematol. 1999 Aug;27(8):1271-81. 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 micro/kappa or micro/lambda 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 DHQ52-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.
本研究比较了人鼠基质细胞支持人类造血干细胞（HSC）向B淋巴细胞系发育的能力。通过荧光激活细胞分选术（FACS）分选的人胎骨髓（BM）HSC（CD34+CD19-或CD34+/CD10-/CD19-/CD45RA）在人胎BM基质细胞、人皮肤成纤维细胞或鼠S17基质细胞上培养，并通过流式细胞术或逆转录聚合酶链反应进行分析。在人BM基质细胞或成纤维细胞上的CD34+CD19- HSC分化为B淋巴细胞系细胞，表面CD19表达密度呈连续变化，且一些细胞表达μ/κ或μ/λ B细胞受体。相比之下，来自S17培养物的CD19+细胞的CD19水平高出两到四倍，但没有细胞表达B细胞受体。在人BM、人成纤维细胞或鼠S17基质细胞培养物中，CD19+细胞的数量和百分比分别为高、中或低。逆转录聚合酶链反应分析表明，当将CD34+/CD19- HSC接种在人或鼠基质细胞上时，末端脱氧核苷酸转移酶（TdT）、CD19和DHQ52-J(H)重排的表达水平相当。相比之下，接种在人或鼠基质细胞上的CD34+/CD10-/CD19-/CD45RA HSC在两种培养物中均表达CD19，但TdT仅在人基质细胞培养物中表达。我们得出结论，人BM基质细胞、人皮肤成纤维细胞和鼠S17基质细胞培养物可为鉴定在调节人类B细胞发育中具有潜在独特功能的基质细胞配体提供互补和比较工具。
