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Research into the effects which normal or immunologically abnormal thymic epithelial cells or epithelial cells of thymoma exert on Blood Stem Cells

正常或免疫异常胸腺上皮细胞或胸腺瘤上皮细胞对造血干细胞影响的研究

基本信息

批准号：
02670611
负责人：
YAMAKAWA Yosuke
金额：
$ 1.54万
依托单位：
Nagoya City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

项目摘要

We made primary culture of thymic tissue obtained from thoracic surgical operation cases. Donors of thymic tissue were classified into four groups. First group is from fetal period to less than 2 year old, second one is from 2 to less than 10, third one is from 10 to less than 20, fourth one is 20 or more.At primary culture, thymic epithelial cells continued to proliferate until end of second week in 1st or 2nd group but in 3rd or 4th group cells barely proliferated after 1 week.Cultured thymic epithelial cells were categorized into three types with shape of cells at primary culture. Type 1 had small cell body and gathered each other. Type 2 had wide body and scattered on the bottom of culture bottle. Type 3 had wide body and gathered each other. Type 1 cells expressed keratin brightly but type 3 cells dully. In type 2 cells, some cells expressed keratin and the others expressed vimentin.We made secondary culture of thymic epithelial cells. At first thymic epithelial cells decreased remarkably in all groups. In 1st or 2nd group, cells started to proliferate at 2nd day of culture but in 3rd or 4th group, cells started to proliferate only a little at 4th day of secondary culture.Mononuclear cells were obtained from bone marrow of ribs at various surgical cases, which were removed cells that expressed CD3 or CD4 or CD8 brightly. Supernatant of thymic epithelial cell culture of each group was added to the CD3-4-8 cell fraction and after 4 days incubation, expression of CD3 or CD4 or CD8 were detected by FACScan. In lymphoid lineage 4th group s ratio was decreased compared with control or 1st or 2nd or 3rd group. In granulocyte lineage 4th one s ratio decreased compared with control. In blast lineage no remarkable change was detected.We analyzed the effects of supernatant of thymic epithelial cell culture. In 1st or 2nd group CD3+4-8- subset and CD3+4+8- subset increased compared with control. In 3rd or 4th group CD3+4+8- subset had no change compared with control.

取胸部手术病人的胸腺组织进行原代培养。胸腺组织供体分为四组。第一组为胎儿期至2岁以下，第二组为2岁至10岁以下，第三组为10岁至20岁以下，第四组为20岁以上。第1、2组胸腺上皮细胞持续增殖至第2周末，第3、4组胸腺上皮细胞1周后几乎不增殖在原代培养时，将细胞分为三种类型。1型细胞体积小，相互聚集。2型体宽，散在瓶底。三阶变异兽体型庞大，彼此之间相互聚集。1型细胞角蛋白表达较强，3型细胞角蛋白表达较弱。在2型细胞中，部分细胞表达角蛋白，部分细胞表达波形蛋白。起初，所有组的胸腺上皮细胞均显著减少。第1组和第2组细胞在培养第2天开始增殖，第3组和第4组细胞在培养第4天开始少量增殖。将各组胸腺上皮细胞培养物的上清液加入到CD 3 -4-8细胞组分中，孵育4天后，通过FACScan检测CD 3或CD 4或CD 8的表达。在淋巴系中，第4组与对照组或第1、2、3组相比，比值降低。粒细胞系第4型细胞比率较对照组降低。我们分析了胸腺上皮细胞培养上清的作用。第1组和第2组CD 3 +4-8-亚群和CD 3 +4+8-亚群较对照组升高。第3、4组CD 3 +4+8-亚群与对照组相比无明显变化。