小地老虎变态期脂肪体变化及保幼激素类似物的影响

本文对小地老虎Agrotis ypsilon(Rottemberg)从四龄幼虫开始经预蛹和蛹的变态期及羽化为成虫后的脂肪体出现的超微结构变化,蛋白质含量的变动,以及蛋白质颗粒的形成和消失过程,进行了系统观察和组织化学分析。结果表明:(1)在幼虫期的后期,脂肪体扩大成宽带状,细胞体积增大的同时出现双核和多核。进入预蛹期,细胞内开始出现嗜碱性“蛋白质颗粒”,血细胞吞噬部分脂肪体细胞。蛹龄一天时,脂肪体转变成块状,细胞内充满大型蛋白质颗粒。在蛹龄5—10天内,“幼虫脂肪体”逐步崩解,围膜及细胞膜消失。至蛹龄12天时转变为预成虫,脂肪体细胞重新出现,并以气管分支为中心聚合成花朵状圆球体,再组成串状“成虫脂肪体”,仍充满蛋白质颗粒。幼虫期发达的粗面内质网和线粒体,至预蛹期则衰变成几种类型的蛋白质颗粒。(2)六龄幼虫在1—5日龄期间,每克脂肪体的蛋白质含量稳定在7.1—8.4毫克之间,随后逐步升高,至预蛹期达16.3毫克的峰值。蛹初期,雄蛹和雌蛹的含量分别增高到预蛹期的1.63和2.4倍。但在蛹龄2—8天内迅速下降到六龄幼虫期的水平。至蛹龄9—13天时间(包括预成虫),含量又突然猛增,雌蛹尤为显著。蛹期脂肪体细胞充满着的几种蛋白质颗粒,在羽化为成虫后的24小时内全部消失。在六龄幼虫期用保幼激素类似物ZR-515(20微克)作体壁处理,可使幼虫期延长4天,并使九日龄幼虫的脂肪体仍保持幼虫型状态。     

小地老虎变态期间马氏管超微结构与酯酶活性的变化

本实验用光镜和电镜观察了小地老虎Agrotis ypsilon Rottemberg幼虫在变态期间马氏管超微结构的变化及成虫马氏管的重组过程,同时还研究了变态期马氏管酯酶的活性.结果表明:(1)变态期间马氏管外形完整,除至预蛹期隐肾复合体解体外,其余无明显变化.(2)变态期间管壁细胞变化显著.幼虫6龄末期马氏管细胞结构开始变化,主要特点为:细胞质电子密度高,充满了核糖体颗粒,微绒毛萎缩,线粒体从萎缩的微绒毛中退出进入细胞质,基膜内褶破坏.进入预蛹期幼虫马氏管细胞解体:基膜内褶、顶端微绒毛、线粒体及细胞质内的其它细胞器消失,并形成自体吞噬泡,细胞质内仅存细胞核及各种类型的液泡.但是在变态期间因底膜始终存在,故马氏管外形不变;至蛹后期,成虫马氏管细胞在原位重组,基膜内褶由浅变深,微绒毛由短变长,线粒体内嵴从无到有.(3)变态过程中羧酸酯酶和酸性磷酸酯酶的活性变化趋势基本相同,以六龄幼虫最强,预蛹期次之,蛹期最低.     

小地老虎变态期间马氏管超微结构与酯酶活性的变化

本实验用光镜和电镜观察了小地老虎Agrotis ypsilon Rottemberg幼虫在变态期间马氏管超微结构的变化及成虫马氏管的重组过程,同时还研究了变态期马氏管酯酶的活性.结果表明:(1)变态期间马氏管外形完整,除至预蛹期隐肾复合体解体外,其余无明显变化.(2)变态期间管壁细胞变化显著.幼虫6龄末期马氏管细胞结构开始变化,主要特点为:细胞质电子密度高,充满了核糖体颗粒,微绒毛萎缩,线粒体从萎缩的微绒毛中退出进入细胞质,基膜内褶破坏.进入预蛹期幼虫马氏管细胞解体:基膜内褶、顶端微绒毛、线粒体及细胞质内的其它细胞器消失,并形成自体吞噬泡,细胞质内仅存细胞核及各种类型的液泡.但是在变态期间因底膜始终存在,故马氏管外形不变;至蛹后期,成虫马氏管细胞在原位重组,基膜内褶由浅变深,微绒毛由短变长,线粒体内嵴从无到有.(3)变态过程中羧酸酯酶和酸性磷酸酯酶的活性变化趋势基本相同,以六龄幼虫最强,预蛹期次之,蛹期最低.     

囊状幼虫病病毒侵染对中华蜜蜂营养和免疫反应的影响

【目的】囊状幼虫病病毒(sacbrood virus, SBV)是严重危害中华蜜蜂Apis cerana cerana蜂群健康和种群数量的病原微生物,但其对蜜蜂的致死机制不明。本研究旨在探究SBV对不同发育阶段中华蜜蜂营养代谢和免疫的影响。【方法】分别给中华蜜蜂2日龄幼虫和新羽化成虫饲喂SBV,逐日统计死亡蜜蜂数量,检测病毒对蜜蜂存活的影响;利用qPCR检测中华蜜蜂4日龄幼虫、预蛹以及10和20日龄成虫体内SBV RNA、营养代谢基因(ilp1, ilp2, hex110, hex70b, hex70c和vg)、先天性病毒免疫基因(rel, toll, apidaecin, abaecin, defensin, hymenoptaecin, jra, key和state92e)、细胞凋亡基因(atg7和LOC100577876)和抗RNA病毒基因(dis3和dicer)的表达水平。【结果】 SBV感染显著降低了中华蜜蜂幼虫的存活率,但对成虫的生存影响不明显。SBV RNA在中华蜜蜂4日龄幼虫和预蛹体内的表达量显著高于其在10和20日龄成虫体内的表达量。SBV显著降低了中华蜜蜂幼虫营养代谢基因ilp1, ilp2, hex110, hex70b和hex70c以及成虫营养代谢基因vg和hex110的表达量,但显著提高了4日龄幼虫rel, toll, apidaecin, abaecin, defensin, hymenoptaecin和jra以及成虫key和 state92e等先天性病毒免疫基因的表达量,还引起预蛹体内的细胞凋亡基因atg7和LOC100577876的表达量显著增加。【结论】SBV在中华蜜蜂幼虫和预蛹体内的感染水平远高于在成虫体内的,其对幼虫的危害也大于对成虫。SBV显著影响了中华蜜蜂正常的营养代谢,染毒中华蜜蜂能够提高自身的免疫水平来应对;预蛹期中华蜜蜂幼虫细胞凋亡水平的显著增加可能与化蛹异常及死亡有关。     

小地老虎蜕皮腺的形态和结构

用电镜和光镜以及组织化学方法研究了小地老虎(Agrotis ypsilon)幼虫蜕皮腺的形态与结构,及其在5龄—6龄幼虫的蜕皮期和预蛹—蛹的变态期的变化,并对蜕皮腺分泌的蜕皮液进行了组织化学分析.结果表明:(1)幼虫期的蜕皮腺共有15对,其中12对位于胸部1—3节和腹部1—9节的背侧面,其余3对位于前、中、后胸基节窝的外侧,至化蛹30小时后逐渐解体;(2)蜕皮腺的分泌细胞均为“液泡型”,在幼虫每次蜕皮期间出现周期性变化,随着蜕皮过程的结束,细胞由分泌状态进入静止状态,体积逐渐变小,色泽由乳白色趋向透明,液泡变小或消失,内质网数量溅少,液泡内含物对PAS反应减弱;(3)蜕皮前蜕皮腺分泌细胞的内含物PAS强阳性,表明蜕皮液是粘多糖或糖蛋白性质的,它通过导管分泌到新旧表皮层之间.     

肠道共生细菌预发酵鸡粪对黑水虻生长发育的影响

以利用黑水虻Hermetia illucens L.肠道共生细菌不同菌株培养液预发酵的鸡粪为培养料,研究了添加不同菌株的发酵鸡粪对黑水虻幼虫生长发育的影响。结果表明,添加不同菌株的预发酵鸡粪能显著增加预蛹和蛹的重量,显著提高化蛹率并增加雌、雄成虫体长,缩短预蛹所需时间,但对幼虫存活率、成虫羽化率、成虫寿命没有显著影响。同时,不同菌株的作用效果有所不同。     

棉铃虫不同发育阶段微粒体P450酶系组成和活性的比较

比较了棉铃虫Helicoverpa armigera 6龄幼虫、蛹、成虫微粒体P450单加氧酶系组成及其活性。P450含量在6龄幼虫中肠>（脂肪体=蛹）>成虫,NADPH-细胞色素还原酶在幼虫中肠>幼虫脂肪体>蛹>成虫;6龄幼虫脂肪体微粒体与蛹脂肪体微粒体P450含量相近,但NADPH-细胞色素还原酶活性前者是后者的4.2倍;成虫微粒体的细胞色素P450和NADPH细胞色素P450还原酶含量很低,几乎未检测出。用对-硝基苯甲醚和艾氏剂为底物测定P450酶系活性表明,与6龄幼虫相比,蛹和成虫具有极低的单加氧酶活性,其O-脱甲基酶活性未检出,艾氏剂环氧化酶活性比幼虫低2～3个数量级。     

几种昆虫病毒交叉感染玉米螟的研究

银纹夜蛾NPV(P_aNPV)、柞蚕NPV(ApNPV)和赤松毛虫CPV(D_aCPV)可感染玉米螟。D_aCPV对玉米螟1龄幼虫的ID_(50)为5.9×10~5PIB/ml饲料,对幼虫的生长发育、蛹重、羽化率和成虫寿命有显著影响。D_aCPV主要侵染幼虫中肠柱状细胞,在细胞质中增殖;P_aNPV和A_pNPV主要侵染幼虫的体壁细胞和气管壁细胞。D_aCPV在玉米螟幼虫体内增殖后,多角体形态由正六角形变为锥形或四方形;成虫羽化时排出的蛹便可观察到多角体,病毒可传递给子代。利用其他昆虫病毒有防治玉米螟的可能性。     

吉林省粘虫猖獗世代发生虫源的研究——Ⅰ.发生虫源的探讨和越冬问题研究

粘虫是吉林省粮食作物的大害虫。本文为作者于1953—1962年研究粘虫的一部分工作,目的在于分析吉林省粘虫发生猖獗世代的虫源问题。我们怀疑春季出现的大量成虫有两种可能来源:(1)本地越冬的成虫、蛹或幼虫;(2)由外地迁入。通过研究分析,所得结果如下: 1.粘虫在吉林省一年完成两个世代。春季发生的第一代是猖獗世代,幼虫为害盛期在6月下旬至7月上旬,其繁殖虫源主要来自5月下旬到6月上半月出现的成虫。 2.粘虫在当地的入冬虫态,有幼虫、蛹及成虫。经过发生地的越冬调查及野外试验,证明粘虫在东北和内蒙地区的自然条件下不论何种虫态,均不能越冬。 3.据粘虫抗寒力测定,在-8℃恒温下,幼虫、蛹、成虫均迅速死亡,在-5℃下最长只能存活3—7小时,在1±1℃下全部死亡的理论时间为成虫19.95天、幼虫44.67天、蛹25.70天。查当地1、2两月旬平均气温最高-9.4℃,最低-16.5℃,田间5厘米深的土温为-12.4—-9.4℃,均超过虫体的死亡低温。 因此,粘虫不可能在吉林省越冬,从而可以推断猖獗世代的虫源,是由外地迁入的。     

黑水虻幼虫的耐饥力及饥饿幼虫复食后的生物学特性

【目的】旨在明确黑水虻Hermetia illucens幼虫的耐饥力及饥饿幼虫复食后的发育、生殖力等生物学特性,为评估饥饿对黑水虻后续发育的影响并指导生产实践提供依据。【方法】将不同日龄或不同体重黑水虻幼虫进行不同时长的饥饿处理,测定和分析其耐饥力与日龄、百头重的关系;对7日龄幼虫进行不同时长的饥饿处理后再复喂,测定不同饥饿时长对其预蛹出现时间、总预蛹率、预蛹重、成虫繁殖等的影响。【结果】黑水虻幼虫饥饿后的半数致死时间(LT₅₀)与日龄成指数关系,与体重呈线性相关。7日龄幼虫的耐饥力较强,LT₅₀超过了50 d,饥饿30 d时存活率仍然达到了95%。百头重小于0.5 g的个体,其LT₅₀小于30 d;百头重在2～2.5 g的个体,其LT₅₀在45～70 d。幼虫超过8日龄时或百头重超过4.5 g时,多数个体在饥饿5 d时就提前进入了后续发育阶段。7日龄幼虫饥饿40 d内复食,其预蛹率未受到影响,但预蛹重显著增加;短期饥饿(0～10 d)对其成虫产卵量无显著影响,长期饥饿(30～40 d)使其成虫产卵量显著下降;饥饿长达30 d以上时,卵孵化率也显著降低。【结论】本研究明确了不同体重或不同日龄的黑水虻幼虫都具有一定的耐饥力;饥饿幼虫复食后预蛹重显著增加。     

Light and electron microscopic studies on the absorptive cells of the intestine, caeca and rectum of the adult rainbow trout, Salmo gairdneri, Rich

The absorptive cells lining the mucosa of the intestine, caeca and rectum of farm-reared adult rainbow trout ( Salmo gairdneri Rich.) were studied by light and electron microscopy. Most of the intestinal absorptive cells showed morphological characteristics of lipid absorption. The cytoplasm of a few cells was closely packed with mitochondria. The caecal absorptive cells resembled those of the intestine but contained more apical dense bodies and more complex lamellar structures which, in conjunction with the intercellular spaces, appeared to play more active roles in lipid absorption. The rectal lumen was divided into peripheral and central parts by the complex mucosal folds. The surface of the peripheral part was mostly lined by vacuolated cells except for small patches of non–vacuolated (generative) cells between the bases of the folds. The central lumen was lined mainly by a second type of non-vacuolated cell. The vacuolated cells showed structural indications of a function of absorption of protein macromolecules.
There is morphological evidence to suggest that the pattern of absorption of lipids and proteins in the adult rainbow trout is similar to that demonstrated in stomachless teleosts and that both intraluminal and intracellular digestion of proteins co-exist in this teleost.     

Ultrastructure of the rectum of infective-stage Wuchereria bancrofti (Nematoda: Filarioidea).

The authors have examined the ultrastructure of the rectum of infective-stage Wuchereria bancrofti by transmission electron microscopy. Our observations show that the rectum is divided into anterior and posterior segments. The cells of the anterior rectum appear to be derived from the microfilarial R (rectal) cells described by other authors. In both stages, these cells show voluminous nuclei, abundant mitochondria, and small cytoplasmic processes which contain fibrillar components. Amorphous material associated with these processes appears throughout the larval rectum and may protrude from the anus as the rectal plug. In the specimens examined, a patent lumen could not be traced completely through the anterior rectum. The posterior rectum has no counterpart in published accounts of microfilarial ultrastructure and probably arises during larval morphogenesis; it is lined with invaginated body cuticle, overlaid by a single layer of epithelial cells which may be of hypodermal origin.     

成体心脏干细胞经静脉移植后在心梗小鼠体内的分布研究

目的：观察心肌梗死小鼠静脉移植成体心脏干细胞后,细胞在小鼠体内各器官的分布情况。明确心梗后静脉移植成体心脏干 细胞在小鼠体内的分布和归巢情况。方法：分离培养小鼠心脏成体干细胞,采用流式细胞仪鉴定细胞,通过亲脂性染料CM-DiI标 记细胞后行小鼠急性心肌梗死模型建立和细胞移植,分别在细胞移植后7、14、8 天取小鼠心脏、肝脏、脑、脊髓、肺脏,行冰冻切 片,在荧光显微镜下观察移植细胞在各组织器官存活和分布情况。结果：成体心脏干细胞分离培养后呈贴壁生长,流式细胞仪检 测显示细胞纯度>80%。CM-DiI标记后荧光显微镜下观察可见标记的细胞胞浆胞核均被染成呈明亮的红色。心肌梗死后经静脉 移植成体心脏干细胞,细胞在各组织中分布呈变化过程,7 天时,在肺脏和肝脏分布较多,至14 天和28 天时,肺脏和肝脏分布减 少,心脏分布逐渐增多,表现出向心脏的" 归巢" 现象,而脑和脊髓在28 天的观察时间内分布较少。结论：采用CM-DiI标记心 脏成体干细胞,操作简单,标记效果好,可用于短期的细胞体内追踪。小鼠心肌梗死后行经静脉成体心脏干细胞移植,28 天后细胞 在心脏的分布逐渐增多,表现出向心脏的" 归巢" 现象。     

Stem cell: balancing aging and cancer

Stem cells are defined by their self-renewing capacity and the ability to differentiate into one or more cell types. Stem cells can be divided, depending on their origin, into embryonic or adult. Embryonic stem cells derive from early stage embryos and can give rise to cells from all three germ layers. Adult stem cells, first identified in hematopoietic tissue, reside in a variety of adult tissues. Under normal physiologic conditions, adult stem cells are capable of differentiating into the limited cell types that comprise the particular tissue or organ. Adult stem cells are responsible for tissue renewal and exhaustion of their replicative capacity may contribute to tissue aging. Loss of unlimited proliferative capacity in some of the adult stem cells and/or their progenitors may have involved the evolutionary trade-off: senescence prevents cancer but may promote aging. Embryonic stem cells exhibit unlimited self-renewal capacity due to the expression of telomerase. Although they possess some cancer cell characteristics, embryonic stem cells exhibit a remarkable resistance to genomic instability and malignant transformation. Understanding the tumor suppressive mechanisms employed by embryonic stem cells may contribute to the development of novel cancer treatments and safe cell-based therapies for age-related diseases.     

Rectal water absorption in seawater-adapted Japanese eel Anguilla japonica

Marine teleosts drink large amounts of seawater to compensate for continuous osmotic water loss. We investigated a possible significant role of the rectum in water absorption in seawater-adapted eel. In rectal sacs filled with balanced salt solution (BSS) and incubated in isotonic BSS, water absorption was greater in seawater-adapted eel than in freshwater eel. Since rectal fluid osmolality was slightly lower than plasma osmolality in seawater-adapted eel, effects of rectal fluid osmolality on water absorption were examined in rectal sacs filled with artificial rectal fluid with different osmolality. Rectal water absorption was greater at lower rectal fluid osmolality, suggesting that an osmotic gradient between the blood and rectal fluid drives the water movement. Ouabain, a specific inhibitor of Na⁺/K⁺-ATPase, inhibited water absorption in rectal sacs, indicating that an osmotic gradient favorable to rectal water absorption was created by ion uptake driven by Na⁺/K⁺-ATPase. Expression levels of aquaporin 1 (AQP1), a water-selective channel, were significantly higher in the rectum than in the anterior and posterior intestines. Immunoreaction for Na⁺/K⁺-ATPase was detected in the mucosal epithelial cells in the rectum with more intense staining in the basal half than in the apical half, whereas AQP1 was located in the apical membrane of Na⁺/K⁺-ATPase-immunoreactive epithelial cells. The rectum is spatially separated from the posterior intestine by a valve structure and from the anus by a sphincter. Such structures allow the rectum to swell as intestinal fluid flows into it, and a concomitant increase in hydrostatic pressure may provide an additional force for rectal water absorption. Our findings indicate that the rectum contributes greatly to high efficiency of intestinal water absorption by simultaneous absorption of ions and water.     

Cryptonephric malpighian tubule system in a dipteran larva,the New Zealand glow-worm, Arachnocampa luminosa (Diptera: Mycetophilidae): A structural study

The Malpighian tubules of the glow-worm are divided into four morphologically distinct regions, each composed of a different cell type. Part 3 of the Malpighian tubules of A. luminosa is intimately bound to the rectum by a layer of fat body. This association of the tubules with the hindgut is referred to as a cryptonephric system. This type of arrangement has been described in some Coleoptera and the larvae of most Lepidoptera but has never before been reported in the Diptera. In the glow-worm the cryptonephric tubules themselves are small, and adjacent to the fat body the epithelial cells are modified to form very thin windows or ‘leptophragmata’ (Lison, 1937). The main epithelial cells exhibit features characteristic of highly active, secretory Malpighian tubule cells. The high density of mitochondria and their association with all the microvilli is indicative of a highly active secretory cell. The high concentration of glycogen in these cells and their intimate association with the hindgut suggest that they may, in addition, have a nutrient absorptive function. The role of the cryptonephric rectal complex in the glow-worm is discussed in the light of present knowledge gained from previous studies of coleopteran and larval lepidopteran cryptonephric systems. On structural grounds a model is proposed for the regulation of the ionic environment of the rectum, and the uptake and metabolism of organic material from the rectal lumen by this cryptonephric complex.     

Adult human CD133/1(+) kidney cells isolated from papilla integrate into developing kidney tubules

Approximately 60,000 patients in the United States are waiting for a kidney transplant due to genetic, immunologic and environmentally caused kidney failure. Adult human renal stem cells could offer opportunities for autologous transplant and repair of damaged organs. Current data suggest that there are multiple progenitor types in the kidney with distinct localizations. In the present study, we characterize cells derived from human kidney papilla and show their capacity for tubulogenesis. In situ, nestin(+) and CD133/1(+) cells were found extensively intercalated between tubular epithelia in the loops of Henle of renal papilla, but not of the cortex. Populations of primary cells from the renal cortex and renal papilla were isolated by enzymatic digestion from human kidneys unsuited for transplant and immuno-enriched for CD133/1(+) cells. Isolated CD133/1(+) papillary cells were positive for nestin, as well as several human embryonic stem cell markers (SSEA4, Nanog, SOX2, and OCT4/POU5F1) and could be triggered to adopt tubular epithelial and neuronal-like phenotypes. Isolated papillary cells exhibited morphologic plasticity upon modulation of culture conditions and inhibition of asymmetric cell division. Labeled papillary cells readily associated with cortical tubular epithelia in co-culture and 3-dimensional collagen gel cultures. Heterologous organ culture demonstrated that CD133/1(+) progenitors from the papilla and cortex became integrated into developing kidney tubules. Tubular epithelia did not participate in tubulogenesis. Human renal papilla harbor cells with the hallmarks of adult kidney stem/progenitor cells that can be amplified and phenotypically modulated in culture while retaining the capacity to form new kidney tubules. This article is part of a Special Issue entitled: Polycystic Kidney Disease.     

Cytological and enzyme-histochemical investigations on the digestive organs of Nautilus pompilius (Cephalopoda, Tetrabranchiata)

The foregut, stomach, caecum, midgut, and rectum of the digestive tract of Nautilus pompilius L.were investigated with ultrastructural and enzyme-cytological methods. Three different cell types were identified within the lamina epithelialis mucosae: main cells, goblet cells, and cells with secretory granules. The main cell type is the epithelial cell with microvilli, a basal nucleus surrounded by dictyosomes, rough endoplasmic reticulum, mitochondria, and electron-dense granules identified as lysosomes in the apical part of the cell. In the caecum this cell type contains endosymbiotic bacteria. The presence of endocytotic vesicles and the storage of lipids in the caecum indicate that this organ is involved in the process of absorption. In the caecum and the longitudinal groove of the rectum the main cells are, in addition, ciliated, facilitating the transport of food particles and faeces. Two types of goblet cells are found in all organs except in the stomach, forming a gliding path for food particles and protecting the epithelium. In the foregut and rectum, cells with electron-dense granules were recognized as the third type. The conspicuous secretory cells of the rectum represent a delimited rectal gland; its possible biological function is discussed. The tunica muscularis in all organs of the digestive tract consists of obliquely striated muscle cells innervated by axons containing transparent, osmiophilic and dense-cored vesicles. Positive reactions for acid and alkaline phosphatase, monoamine oxidase, β-glucuronidase, and trypsin- and chymotrypsin-like enzymes are localized in the lamina epithelialis mucosae.     

Ultrastructure of the Rectum Epithelial Cells in the Mosquito Larvae, Anopheles sinensis Wiedemann

ABSTRACT The Ultrastructure of rectum epithelial cells in the mosquito larvae, Anopheles sinensis Wiedemann, was studied using electron microscope. The rectal epithelium forms rectal papillae composed of the absorptive cells and the surrounding basal cells. Moreover, rectal epithelium was covered with thin cuticular intima. Apical plasma membrane of the epithelial cells had infoldings and in between them, mitochondria developed into elongated shape were attached. In addition, the membrane infoldings reach down into the cell cytoplasm to form several layers of leaflet-like prolongations. On both sides of these prolongations were also large, well-developed mitochondria. Their formation was that mitochondria were attached to 3 μm length and 4–13 layers of membrane wrinkle lump. Many spherites, which are lamelated crystals that form an illusory structure in concentric circles inside of the cytoplasm of epithelial cell were observed. Basal plasma membrane in the epithelial cells was also wrinkled to promulgate into the cytoplasm to become basal infoldings producing canaliculi in basal labyrinth formation. There were many mitochondria scattered in these formations as well. On the bottom of the epithelial cell, basal lamina was attached and between basal lamina and muscle bundle was subepithelial space, which is connective tissue. Inside the space, tracheal and nerve cells were observed.     

成人宫颈上皮细胞的原代培养及鉴定

目的:探索采用无血清培养基原代培养成人宫颈上皮细胞的方法。方法:以成人的宫颈上皮组织为研究对象,采用胰蛋白酶-EDTA消化法获得宫颈上皮细胞悬液,于上皮细胞专用无血清培养基中培养,采用免疫细胞化学法测定细胞中角蛋白及波形蛋白的表达,对细胞纯度进行鉴定。结果:原代培养10-15天细胞融合达60%,传代至4-6代,细胞出现生长衰退。早期细胞生长状态良好,细胞纯度在90%以上。结论:采用酶消化法及K-SFM无血清培养基培养可获得纯度高的成人宫颈上皮细胞。