人参皂甙对小鼠腹水型网状细胞肉瘤ARS细胞分化与生长的影响

用人参皂甙怍用于短期体外培养的 ARS 细胞,取盖片或搜集瘤细胞,油镜下测量200个瘤细胞、核及核仁的直径(μm)。对照组:细胞9.77;核6.57;核仁2.96;核质比值0.68。实验组:细胞9.24;核5.61;核仁2.75;核质比值0.61。核仁 P<0.01,其余 P<0.001。实验组瘤细胞、核、核仁及核质比值均明显减小。核质比值小于0.5的小细胞增多,核仁亦减小的小细胞,形态近似巨噬细胞。扫描电镜观察见实验组小细胞增多,表面微绒毛减少,出现皱褶。透射电镜观察见实验组瘤细胞内细胞器增多;线粒体变小,数量增多,大小一致,分布规则,均提示细胞趋向分化。实验组瘤细胞内非特异性酯酶和颗粒增多;吞噬 Polystyrene胶乳颗粒的细胞,由对照组的77.5%增至98.5%;     

dbcAMP对小鼠U_(14)腹水型瘤细胞的影响——扫描电镜观察和流式细胞光度计分析

本文根据扫描电镜观察,报告了dbcAMP引起小鼠U_(14)腹水瘤细胞微绒毛和质膜表面的细微形态变化。U_(14)腹水瘤的多数细胞体积大,大小相近,表面有许多细长微绒毛;微绒毛表面和微绒毛间细胞质膜表面,仅见很细微的点状微突。腹腔内注射dbcAMP的U_(14)腹水瘤,85％以上的细胞体积缩小,大小不等,表面微绒毛缩短且减少。在没有微绒毛的部位,细胞质膜表面有许多颗粒状微突起,较对照组明显易见。微绒毛表面也有与之大小类似的微突起,使微绒毛表面凹凸不平。流式细胞光度计分析结果表明,在本文实验条件下,dbcAMP未引起U_(14)腹水瘤细胞周期的变化,细胞膜表面和微绒毛的变化,也不单纯是细胞周期中微绒毛数量和分布的改变。因此作者认为,这些变化不是通过改变细胞周期引起的。 cAMP引起微绒毛退缩的机制不详。至于微绒毛间的颗粒状微突起是微绒毛退缩的遗迹,抑或是由cAMP引起,尚待研究。cAMP引起瘤细胞表面形态的变化,进一步证明了cAMP在小鼠体内诱导恶性细胞分化的作用。     

人早幼粒白血病细胞与小鼠网织红细胞融合的胞质杂交细胞形态学观察

本研究对HGPRT缺失的人早幼粒白血病细胞突变株(HL-60-AR)与小鼠网织红细胞融合所形成的胞质杂交细胞,进行了光镜和电镜的形态学观察。所得结果显示:胞质杂交细胞在短期培养过程中,可见大量细胞呈现核固缩和核偏位,甚至出现排核现象;长期培养的胞质杂交细胞沿不同途径向较成熟的方向分化。本文并讨论了网织红细胞胞质因子对人早幼粒白血病细胞分化途径的影响。     

诱导大鼠间充质干细胞形成的神经元样细胞的形态特征

目的 研究诱导大鼠骨髓间充质于细胞(MSCs)形成的神经元样细胞的形态特征.方法 通过贴壁法培养鉴定大鼠骨髓MSCs,体外培养扩增纯化后加入新生鼠脑匀浆上清诱导48 h.倒置显微镜和透射电镜下观察诱导前后细胞的形态结构变化,免疫细胞化学方法鉴定诱导后细胞的性质.结果 倒置显微镜下可见MSCs呈纺锤形和多角形,核居中,有1或2个核仁,诱导后细胞呈神经元样,细胞伸出较长的轴突样和树突样突起;免疫组织化学方法显示NSE、NF阳性,GFAP阴性.透射电镜下可见诱导前MSCs胞质较丰富,有较多的细胞器,细胞核呈圆形或椭圆形,也可见不规则形核,核染色质较疏松,有明显核仁,多为1个.有的MSCs表面可见较多的微绒毛.诱导后细胞胞质丰富,细胞核多为不规则形,染色质疏松,核仁较大,1～3个不等,细胞表面有发达的微绒毛.结论 MSCs是一种多分化潜能的细胞,诱导后分化成为具有成熟细胞器的神经元.     

观察佛波脂和二甲基亚砜联合诱导人肺腺癌GLC分化过程中的形态和超微结构

目的 探讨佛波脂(TPA)和二甲基亚砜(DMSO)联合诱导分化剂对人肺腺癌GLC细胞形态的影响。方法 采用细胞计数，光镜、扫描电镜与透射电镜观察，经过TPA和DMSO联合诱导的人肺腺癌GLC细胞增殖以及形态和超微结构的变化。结果 经过联合诱导分化剂作用后，细胞增殖受到抑制，细胞体积增大，呈现扁平铺展状态，细胞核质比例变小，核仁数量减少。细胞表面微绒毛减少，细胞边缘丝状伪足减少，片状伪足增多，核形态较规则，核内异染色质减少，常染色质增多，细胞质中的细胞器数量增多，结构趋于正常。结论 TPA和DMSO联合诱导能够一定程度改变GLC细胞恶性形态结构特征并对GLC细胞具有一定的诱导分化作用。     

含白细胞介素-3上清液促进小鼠骨髓基质细胞生长作用的体外观察

在培养液中加入WEHI-3上清液(含白细胞介素-3,IL-3)作为细胞刺激因子培养小鼠骨髓细胞,第6d培养的细胞结果表明,WEHI-3上清液能在体外促进基质细胞贴壁增殖。含20%WEHI-3上清液的实验组细胞数量为1.334×10~6个/L,光镜和扫描电镜下可见各具特征的4种基质细胞:1.类成纤维细胞:胞体呈长梭形,表面有许多丝状和微绒毛样的突起;2.网状细胞:胞体不规则,有树突状突起,偶见有沿突起走行的皱褶;3.类巨噬细胞:胞体椭圆形,细胞内含吞噬颗粒,ANAE阳性,表面有片层样皱褶和长丝状突起;4.脂肪细胞:此类细胞较少见,胞体呈圆形,细胞内含脂质,苏丹黑B染色阳性,表面平滑。同时,光镜和扫描电镜下均可见基质细胞与造血细胞间有密切接触。而对照组贴壁细胞数少,细胞胞体小,且胞质较少,酶活性也差。细胞数仅为58.83/mm~2。扫描电镜下观察,细胞胞体较小,表面低平。本研究结果表明,WEHI-3上清液具有促进小鼠基质细胞生长和增殖的作用,并可引起相应的形态及组织化学改变。     

正常小鼠小肠微皱褶细胞的形态学观察

目的 观察正常小鼠小肠Peyer‘s集合淋巴小结滤泡相关上皮中微皱褶细胞的形态结构。分布及其表面特性。方法 常规的扫描电镜,透射电镜和免疫荧光,免疫冷冻超薄切片技术,结果 微皱褶细胞的粘膜面具有许多短而不规则的微绒毛和微皱褶,基部胞膜向顶部呈穹隆状突起,形成“口袋”样结构,其内含有B、T淋巴细胞和少量的巨噬细胞,顶部胞质内终末网不发达,有许多小泡;荆豆凝集素UEA-1可特异性地与微皱褶细胞结合。结论 微皱褶细胞是分布于Peyer‘s集合淋巴小结圆顶区表面滤泡相关上皮中的一种特化的上皮细胞,利用标记的凝集素UEA-1可作为鉴别微皱褶细胞的依据。     

三七皂甙R_1对HL—60细胞系体外诱导分化作用的初步研究

作者经光镜及电镜形态观察发现,用三七皂甙R_180μg/ml处理5日,可诱导68％的HL—60细胞分化,其中晚幼粒32％,杆状30％,分叶6％。经药物诱导前HL—60细胞为圆形或不规则椭圆形,核大而圆,胞浆少而染色浅,95％细胞呈原粒及早幼粒形状;经药物诱导后HL—60细胞胞体明显变小,核质比例降低,核形状不规则,出现肾形杆状及分叶核。首次证实三     

大鼠肾脏间质细胞增龄性变化的透射电镜观察

本文观察了二组不同年龄大鼠肾间质细胞的超微结构,结果表明,肾间质细胞有增龄变化,主要包括:细胞数量、胞质、胞质突起、细胞器增多,核周池缩小。     

小鼠类巨噬细胞系超微结构的观察

本文报道了小鼠类巨噬细胞系(MMC-1)的超微结构。在透射电镜下,可见暗细胞、亮细胞及空细胞。这三种细胞有一定的比例。在其他培养细胞中也见到了类似的三种细胞。暗细胞电子致密度最高,呈圆形、椭圆形或梭形。表面有许多突起,有的呈丘状。胞质可分内、外两部分。外胞质无细胞器;内胞质中有丰富的线粒体、粗面内质网和聚核蛋白体。溶酶体易见,高尔基复合器发达,有成堆的微纤维。胞核不规则,多呈肾形,位于细胞的一侧。核膜清楚,常染色质较多。有的胞核中可见核仁。亮细胞突起少,线粒体肿胀,溶酶体与脂滴易见,核肿胀。空细胞的突起更少,内质网明显减少或消失,溶酶体多,次级溶酶体易见胞核大,染色质稀疏。本文根据此细胞的超微结构特点,证实其为巨噬细胞,并根据其来源称之为类巨噬细胞。     

Blast-like cell compartment in carcinogen-induced proliferating bile ductules.

Small non-epithelial cells with morphological features of blast-like cells are found within a proliferating intrahepatic biliary system after institution in rats of a diethylnitrosamine, 2-acetylaminofluorene, partial hepatectomy carcinogenesis protocol. Two to three days after the partial hepatectomy step of the carcinogen protocol, the small blast-like cells are evident beneath a layer of bile ductule epithelial cells that line the walls of the bile ductules. The basally located small cells are not exposed to the bile ductule lumen or to the surrounding basal lamina. They ranged in size from 3.0 to 5.0 microns, exhibit an undifferentiated phenotype, including a high nucleus-to-cytoplasm ratio and no to minimal differentiated cytoplasmic and surface structures. Mitosis of blast-like cells are evident, and their nuclei express proliferating nuclear cell antigen. The ductal blast-like cells do not express cytokeratin 19, oval cell antigen 270.38, or actin immunoreactivity, in contrast to bile ductule epithelial cells. The basal cells, as well as bile ductule epithelial cells, are negative for a panel of T and B lymphocyte surface markers in contrast to lymphocytes present in the connective tissue stroma surrounding the bile ductules and throughout the hepatic parenchyma. Within some segments of the biliary system, some of the ductal blast-like cells increased in size to approximately 10 microns and showed increased amounts of cytoplasmic organelles and plasma membrane filapodia but did not develop the polarized phenotype of bile ductule epithelial cells (ie, apical microvilli, desmosomes, connections to bile ductule cells, and exposure to duct lumen); however, their nuclear morphology was essentially similar to the smaller basal cells. We also found bile ductules to contain two types of polarized epithelial cells, one with the characteristic oval nucleus of the oval/bile ductule epithelial cells and the other, transitional epithelial cells with a rounder nucleus and prominent nucleoli. The transitional cells exhibit a similar apical-basal polarity and antigenic phenotype as the oval/bile ductule epithelial cells. However, transitional cells are larger and have an overall less dense cytoplasm than the bile ductule epithelial/oval cells, and some show apical microvilli changes and small catalase-positive peroxisomes. These observations indicate that a greater diversity of cell types exist within intrahepatic bile ductules of rats treated with carcinogens. Furthermore, the nonpolarized ductal blast-like cells undergo proliferation and are significantly different in phenotype from other hepatic cells previously reported as candidates for liver progenitor cells.     

猫、兔、鼠侧脑室脉络丛的扫描电镜观察

本文报道用扫描电镜观察猫、兔和大白鼠侧脑室脉络丛的微细结构。3种动物侧脑室脉络丛表面均可见密集的微绒毛、散在的纤毛簇、单个的纤毛、球样突起和花样结构。在高倍镜下,微绒毛又可分为两型,即尖细的指样绒毛和末端膨大成泡样的微绒毛。并发现有呈花样密集排列的微绒毛簇。同时在3种动物的侧脑室脉络丛表面均可见kolmer细胞,根据突起的多少可将此类细胞分成为单极、双极,多极、无伪足样突起四型。猫的侧脑室脉络丛的球样突起和大白鼠的纤毛簇较多。     

人胎儿脑室系统接触脑脊液神经元的扫描电镜观察

用扫描电镜较全面地观察了人胎儿脑室系统－侧脑室、第三脑室和第四脑室壁的超微结构。结果发现人和某些动物一样，几个脑室室管膜表面都覆盖着大量的纤毛和微绒毛。纤 的分布在区域上有一定的差别。并证实了３个脑室内存在着接触脑脊液神经元的胞体、树突和轴突该神经元的胞体为梭形或球形，可见到一个或两个以上的突起。室管膜上神经纤维发自神经细胞或自室腔外穿入而来。另外，在室管膜下还观察到了神经胶质样细胞和类组织细胞。     

Scanning and transmission electron microscopic study of visceral and parietal peritoneal regions in the rat

The visceral peritoneum of intraabdominal organs (spleen, stomach, liver, small intestine), omentum majus and the parietal peritoneum of the anterior abdominal wall and the diaphragm were studied in adult Wistar rats by combined scanning and transmission electron microscopy (SEM, TEM). In general, the peritoneal surface consisted of a mesothelium composed of cubic, flat or intermediate cell types delimited by a basal lamina. Cubic mesothelial cells predominated in parenchymal organs (spleen, liver) and were characterized by prominent and indentated nuclei, a cytoplasm richly supplied with organelles, a dense microvillous coat, basal invaginations and elaborate intercellular contacts. Flat mesothelial cells were observed in the intestinal, omental and parietal peritoneum (tendinous diaphragm, abdominal wall) and showed elongated nuclei, scant cytoplasm, a poorly developed organelle apparatus and sparsely distributed microvilli. An intermediate mesothelial cell type was described within the gastric peritoneum characterized by a central cytoplasmic protrusion at the nuclear region containing most of the cytoplasmic organelles and by thin finger-like cytoplasmic processes. The submesothelial connective tissue layer was composed of collagen fiber bundles, fibroblasts and free cells (macrophages, granulocytes, mast cells) and contained blood and lymphatic vessels. In the spleen, elastic fibers formed a membranous structure with intercalated smooth muscle cells. Mesothelial openings were observed as tunnel-like invaginations within the hepatic peritoneum and as clusters of peritoneal stomata within the parietal peritoneum of the anterior abdominal wall and the muscular diaphragm. The round or oval openings of the peritoneal stomata were frequently occluded by overlapping adjacent mesothelial cells and their microvillous coat or obstructed by cellular material. At the side of the peritoneal stomata the mesothelial cell layer was interrupted to allow a direct access to the underlying submesothelial lymphatic system. The mesothelium and lymphatic endothelium shared a common basal lamina. The endothelial cells were discontinuous and displayed valve-like plasmalemmatic interdigitations facilitating an intercellular transport of fluids and corpuscular elements from the peritoneal cavity to the submesothelial lymphatic lacunae. The findings underline the morphological heterogeneity of the peritoneum in visceral and parietal regions, suggesting different functional implications, and further support the presence of extra-diaphragmatic peritoneal stomata.     

Ultra-high-resolution scanning electron microscopic studies on the membrane system of the parietal cells of the rat in the resting state and shortly after stimulation

The three-dimensional organization of the membrane system of the rat parietal cells in the resting state and during early stimulation with tetragastrin (gastrin) was determined by ultra-high-resolution scanning electron microscopy. Specimens were prepared by cytoplasmic matrix removal using the aldehyde-osmium-DMSO-osmium procedure. The intracellular canaliculus was lined with numerous microvilli. Viewed from the cytoplasmic side, the intracellular canaliculi appeared as an arborized system of cactus-like structures with numerous round holes about 100 nm in diameter corresponding to the basal openings of the microvilli. The intracellular canaliculi were more developed after gastrin stimulation than in the resting state. In resting cells, most of the tubulovesicles were isolated, 100–200 nm in diameter, spherical or tubular in shape, and had a smooth surface. After gastrin stimulation, these structures were interconnected by slender tubules of about 30 nm in diameter forming together tubulovesicular network. Occasionally, swollen and shrunken profiles were observed. The tubulovesicular network was connected with the intracellular canaliculus only at a few sites by the slender connecting tubules. Fusion of the tubulovesicular network with the intracellular canaliculus is observed at such sites. In the fasted rat, the microvilli were slender and their interior was packed with some kind of ill-defined material, probably microfilaments. However, after gastrin stimulation, the microvilli were swollen and their interior was almost empty. These morphological changes seem to indicate the accumulation of fluid in the microvilli after gastrin stimulation, with subsequent swelling. © 1993 Wiley-Liss Inc.     

Electron microscopic observations in in situ and microinvasive bronchogenic squamous cell carcinoma

Seventeen cases of resected in situ and microinvasive bronchogenic squamous cell carcinoma were studied by light and electron microscopy. No definite secretory differentiation was found in any case. Examination of the tumour cells in the basal layer for electron density of cytoplasm, intercellular spaces, and degree of development of cytoplasmic processes showed a variety of cells ranging from type I, where the cytoplasm was dark, development of cytoplasmic processes was good, and the intercellular spaces were large, to type III, where cytoplasmic processes and intercellular spaces were less well developed and the cells were mostly of clear cell type. The tendency to invasion was greater in type III than type I and there was also more marked cellular atypia, more extensive dissolution of basement membrane, a larger number of mitotic figures per 3000 cells in the basal layer, and greater enlargement of nuclear and cytoplasmic areas. A good rank correlation coefficient was obtained. Small dense-core granules were observed in some cases. These finding suggest the strong possibility that cell kinetics and cellular morphology are related to the development of squamous cell carcinoma.     

Differentiation of a colon cancer cell line on a reconstituted basement membrane in vitro

Basement membrane, a thin extracellular matrix, functions as a tissue stabilizer that promotes tissue integrity and differentiated phenotype. We studied a human colon cancer cell line, SNU 61, to evaluate its ability to differentiate on basement membrane. Cells were cultured on plastic, reconstituted basement membrane (Matrigel) or polyhydroxyethyl methacrylate (poly HEMA) for 72 h and evaluated by light and electron microscopy. On Matrigel, the cells showed gland formation with highly polarized cells containing basal nuclei and well developed brush border microvilli on the luminal surface. Apoptosis was noted mainly at the luminal side. On electron microscopic examination, numerous long microvilli, abundant cytoplasmic organelles and intercellular junctions were noted in the Matrigel-cultured cells. Intermediate cytoskeletons were scattered in the cytoplasm and existed on the axes of microvilli. Junctional complexes and desmosomes were frequently formed along intercellular spaces. The cells cultured on poly HEMA, on the other hand, were poorly differentiated and contained a few glandular structures with small lumens. Brush border microvilli, characteristic of enterocytic differentiation, were few in number and were developed on the basal surface. Intermediate filaments and microtubules were fewer than in the Matrigel-cultured cells. Carcinoembryonic antigen was expressed on the luminal surface of the Matrigel-cultured cells and in the cytoplasm of the poly HEMA cultured cells. CD44 stained the basolateral surface in the Matrigel-cultured cells, but the basal side was not stained in the poly HEMA cultured cells. These results are consistent with the different localization of microvilli in the Matrigel and in the poly HEMA cultured cells. Our observations suggest that human colon cancer cells on basement membrane can undergo glandular differentiation and that extracellular matrix is an important factor in morphogenesis.     

Ependymoma: Ultrastructural Studies of Two Cases

Several unusual ultrastructural findings in two ependymomas are reported. In case 1, a grade I ependymoma of the fourth ventricle, there were rosettes, perivascular pseudorosettes, and tumor cells having unusual intracytoplasmic vacuoles by light microscopy. Ultrastructurally, these vacuoles were frequently microrosettes as well as scattered, degenerated cytoplasmic processes of tumor cells. The lumina of some of the microrosettes were bordered by abnormally long and malformed zonulae adherentiae. In case 2, a recurrent grade III ependymoma of the third ventricle, there were rosettes and perivascular pseudorosettes as well as more cellular and anaplastic areas by light microscopy. Ultrastructurally, the cytoplasmic processes of tumor cells in perivascular pseudorosettes contained frequent dense-core vesicles and occasional parallel arrays of microtubules. These structures do not occur in normal mammalian ependymal cells but do occur in the ependymal tanycyte, a related cell that is plentiful in the walls of the third ventricle. Thus some of the tumor cells of this third ventricle ependymoma appear to have differentiated as tany-cytes.     

Ependymoma: Ultrastructural Studies of Two Cases

Several unusual ultrastructural findings in two ependymomas are reported. In case 1, a grade I ependymoma of the fourth ventricle, there were rosettes, perivascular pseudorosettes, and tumor cells having unusual intracytoplasmic vacuoles by light microscopy. Ultrastructurally, these vacuoles were frequently microrosettes as well as scattered, degenerated cytoplasmic processes of tumor cells. The lumina of some of the microrosettes were bordered by abnormally long and malformed zonulae adherentiae. In case 2, a recurrent grade III ependymoma of the third ventricle, there were rosettes and perivascular pseudorosettes as well as more cellular and anaplastic areas by light microscopy. Ultrastructurally, the cytoplasmic processes of tumor cells in perivascular pseudorosettes contained frequent dense-core vesicles and occasional parallel arrays of microtubules. These structures do not occur in normal mammalian ependymal cells but do occur in the ependymal tanycyte, a related cell that is plentiful in the walls of the third ventricle. Thus some of the tumor cells of this third ventricle ependymoma appear to have differentiated as tany-cytes.     

Abnormal nuclear structures (micronuclei, nucleoplasmic bridges, and nuclear buds) in a pleomorphic giant cell carcinoma of the stomach

A rare case of pleomorphic giant cell carcinoma of the stomach in a 70-year-old man is reported. Characteristic microscopic findings included a general lack of architectural cohesiveness, aggregates of mononucleated or multinucleated giant cells, extensive areas of coagulative necrosis, and numerous mitoses. Immunohistochemically, tumor cells displayed cytoplasmic immunoreactivity for cytokeratin AE1/AE3 as well as overexpression of p53 and Ki-67. Electron microscopy revealed paranuclear tonofilaments bundles in giant cells confirming their epithelial nature. Furthermore, giant cells contained two or more nuclei with heterogeneous size, nucleoplasmic bridges, nuclear buds, and micronuclei. Similar abnormal nuclear structures have been closely related to breakage-fusion-bridge type of mitotic disturbances in tumor cell lines, and have not been previously reported in a human tumor.