Pietrangeli C E, Hayashi S, Kincade P W
Oklahoma Medical Research Foundation, Oklahoma City 73104.
Eur J Immunol. 1988 Jun;18(6):863-72. doi: 10.1002/eji.1830180606.
Stromal cells which grow as an adherent layer of Whitlock-Witte cultures are thought to be an essential component of the lymphohemopoietic microenvironment. Stromal cell lines from bone marrow (BM) and spleen have been obtained by treatment of cultures with 5-fluorouracil and selected for their lymphocyte support capacity by measuring the clonal growth of stromal cell-dependent lymphocyte lines in methyl cellulose. Established stromal cell lines differed significantly from stromal cells in primary Whitlock-Witte cultures with respect to expression of certain hemopoietic cell surface markers. For example, the Thy-1 and Mac-3 antigens were expressed by stromal cell lines obtained from BM and spleen, but not by stromal cells in primary cultures. Features common to all stromal cells include synthesis of actins, the neural adhesion molecule N-CAM, and a variety of collagens. Two types of common leukocyte antigens were not significantly expressed. The proliferation and total protein synthetic capacity of lymphocyte-supportive stromal cell lines was sensitive to ionizing radiation. After exposure of the cells to 200 rads, the incorporation of either [3H]thymidine or [3H]Leucine was reduced to less than 50% of control values, but the growth of lymphocytes was augmented in the presence of an irradiated stromal cell layer. The proliferation of stromal cell lines was also affected by exposure to a variety of growth factors. Addition of epidermal growth factor or endothelial cell growth factor augmented BM or spleen-derived stromal cell proliferation, while interferon-gamma had the opposite effect. In general, but not exclusively, lymphocyte growth was inhibited by factors which augmented the proliferation of stromal cells. Novel methods are described for isolating stromal cells and determining their capacity to support lymphocyte growth in vitro. Evidence is presented that this ability is not restricted to BM-derived stromal cells. The function of stromal cells was not dependent on their ability to proliferate, and this may be modulated by immunoregulatory and other growth factors.
作为惠特洛克 - 维特培养物贴壁层生长的基质细胞被认为是淋巴细胞生成微环境的重要组成部分。通过用5 - 氟尿嘧啶处理培养物获得了来自骨髓（BM）和脾脏的基质细胞系，并通过测量甲基纤维素中基质细胞依赖性淋巴细胞系的克隆生长来选择其淋巴细胞支持能力。已建立的基质细胞系在某些造血细胞表面标志物的表达方面与原代惠特洛克 - 维特培养物中的基质细胞有显著差异。例如，从BM和脾脏获得的基质细胞系表达Thy - 1和Mac - 3抗原，但原代培养物中的基质细胞不表达。所有基质细胞共有的特征包括肌动蛋白、神经粘附分子N - CAM和多种胶原蛋白的合成。两种常见的白细胞抗原未显著表达。支持淋巴细胞的基质细胞系的增殖和总蛋白合成能力对电离辐射敏感。细胞暴露于200拉德后，[3H]胸腺嘧啶或[3H]亮氨酸的掺入量降至对照值的50%以下，但在有辐照的基质细胞层存在的情况下淋巴细胞的生长增强。基质细胞系的增殖也受到多种生长因子暴露的影响。添加表皮生长因子或内皮细胞生长因子可增强BM或脾脏来源的基质细胞增殖，而干扰素 - γ则有相反的作用。一般而言，但并非唯一，增强基质细胞增殖的因子会抑制淋巴细胞生长。描述了分离基质细胞并确定其体外支持淋巴细胞生长能力的新方法。有证据表明这种能力不限于BM来源的基质细胞。基质细胞的功能不依赖于其增殖能力，并且这可能受到免疫调节和其他生长因子的调节。
