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张家骅 《国外兽医学(畜禽疾病)》1995,16(4):6-9
哺乳动物睾丸的内分泌功能和生精功能主要由促性腺激素控制，但睾丸内局部因子的自分泌和旁分泌作用起着重要的调节作用，本文着重介绍支持细胞与间质细胞、支持细胞与管周细胞，支持细胞与管周细胞，支持细胞与生殖细胞之间各种因子的调节作用和相互关系。 相似文献
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为研究促进精子发生的体外培养体系,通过无血清睾丸组织培养法,将成年牛睾丸组织块种植于培养板, 观测不同时间睾丸细胞的迁出和形态变化。通过油红O染色鉴定具有分泌功能的支持细胞,通过碱性磷酸酶和免疫组化染色鉴定集落样生长的精原干细胞。结果发现该体系可获得生长良好的多种类型细胞,包括表达AP、Oct-4和GFRα1的精原干细胞及具有分泌功能的睾丸支持细胞,并可获得大量精子。说明,该无血清培养体系可作为一个重要的工具来研究生物学和化学因素对雄性生殖系统的影响,也为研究精子体外发生和移植提供了材料。 相似文献
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旨在建立牦牛与犏牛睾丸支持细胞分离培养及鉴定方案,比较两种牛睾丸支持细胞的生物学特性。本研究分别采集24月龄的3头健康公牦牛与3头F1代公犏牛睾丸组织作为2个样品组,其中每组3个生物学重复,分别通过混合酶消化、差速贴壁和饥饿处理分离得到两种牛的睾丸支持细胞,采用DMEM高糖及DMEM/F12培养基培养睾丸支持细胞,筛选更佳的培养体系。采用碱性磷酸酶染色、油红O染色和免疫荧光染色鉴定细胞表型特征,利用CCK8和RT-qPCR法分别检测细胞的增殖活性和标志功能基因的表达,进一步通过不同浓度丝裂霉素C处理两类支持细胞来评价两牛种支持细胞的耐受性及其作为饲养层细胞的潜能。结果,经形态、特殊染色及标志基因表达鉴定,成功分离到牦牛与犏牛睾丸支持细胞,建立了牦牛与犏牛睾丸支持细胞体外长期培养方案。发现DMEM高糖培养基更适用于睾丸支持细胞的增殖,两牛种支持细胞形态相似、轮廓清晰、呈现多边形或长梭形,牦牛睾丸支持细胞的体外增殖速率及活性远高于犏牛。调节精原细胞增殖分化相关基因（胶质细胞源性神经营养因子（glial cell line derived neurotrophic factor,GDNF）和转化生长因子β1基因（transforming growth factor-β1,TGF-β1））的表达在犏牛支持细胞中分别下调了3.4与2.9倍（P<0.05）,基质细胞源性因子12基因（stromal cell-derived factor 12,CXCL12）的表达在犏牛上调了3.6倍（P<0.05）;调节睾丸发育的SRY-盒包含蛋白9基因（sex determining region Y-box9,SOX9）和睾丸支持细胞特异表达的Wilm肿瘤基因1（Wilms tumor gene1,WT1）在犏牛支持细胞中的表达分别下调了25.9（P<0.01）与38.7倍（P<0.01）。与牦牛相比,犏牛睾丸支持细胞对于丝裂霉素C具有较差的耐受性,表现为细胞核质分界不清、胞质空泡化严重和悬浮死细胞增多。本研究成功建立了牦牛与犏牛睾丸支持细胞分离纯化与体外培养方案;与牦牛相比,犏牛睾丸支持细胞在增殖活性和睾丸生精细胞分化关键功能基因表达等方面均存在缺陷,这可能也是导致犏牛雄性不育的原因之一。 相似文献
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支持细胞也称足细胞,是由早期发育着的胚胎生殖脊细胞分化而成,它与精原细胞一起构成睾丸曲细精管。支持细胞具有营养生精细胞的作用,生精细胞发育过程中所需的营养物质的摄入及代谢废物的排出,都必须通过支持细胞的转运。支持细胞能分泌多种物质,其中分泌的蛋白类主要有转运蛋白和调节蛋白两种。 相似文献
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贵州香猪睾丸发育中支持细胞和生精细胞数量变化观察 总被引：1，自引：0，他引：1
为了解香猪睾丸发育过程中生殖细胞和支持细胞数变化规律,用手术取出30,40,50,70,90和110日龄(每个年龄组n=3～4)香猪右侧睾丸,经中性多聚甲醛固定,石蜡包埋,组织切片采用免疫组化SP法,用单克隆抗体GATA-4检测睾丸支持细胞的特异生长转录因子-4,经DAB显色、苏木素复染。光镜下核呈棕色者为支持细胞,核呈蓝色者则为生殖细胞;经显微照相并用Scion image软件测量生精小管及管壁面积。结果:30～110日龄睾丸支持细胞数维持在稳定水平(P>0.05),而生殖细胞数随日龄增加而增多,70日龄生殖细胞数快速增多(P<0.05),持续到110日龄。同样,从70日龄开始睾丸生精小管和管壁面积显著性增大(P<0.05)。香猪睾丸支持细胞快速增殖发生在30日龄前,而生殖细胞数随着日龄的增长而增多。 相似文献
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《中国兽医杂志》2016,(6)
为了观察小鼠氟中毒睾丸形态变化特点,研究氟对睾丸的毒性影响,选用45只健康性成熟雄性昆明系小鼠,随机分为对照组、低氟组(25 mg/L Na F)、高氟组(100 mg/L Na F),连续饲喂60 d,制作睾丸石蜡组织切片,H.E.染色后,显微镜下观察曲细精管形态变化,通过定量分析方法探讨氟对睾丸的毒性影响。结果表明,与对照组相比,低氟组和高氟组曲细精管半径、面积以及曲细精管管腔半径、面积均无显著性差异(P0.05);高氟组小鼠曲细精管细胞层数和厚度极明显低于对照组(P0.01);形态学观察显示,25 mg/L氟中毒小鼠睾丸生精细胞减少,100 mg/L Na F可导致生精细胞排列紊乱、各级生精细胞减少等。本试验可为氟的雄性生殖毒性研究提供直观、科学的参考。 相似文献
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鸡胚睾丸支持细胞的传代培养与鉴定 总被引：1，自引：0，他引：1
以孵化18d的广西土鸡鸡胚为实验材料,采用两步酶消化法分离鸡胚睾丸支持细胞,置于37℃、5％C02的培养箱中培养,用低渗的磷酸缓冲液（PBS）处理进行纯化,并对原代和传代支持细胞进行了鉴定。结果显示,睾丸支持细胞生长良好,已传至第4代;经低渗处理,可去除绝大部分的生精细胞,获得的支持细胞纯度可达95％;鉴定结果显示,支持细胞为碱性磷酸酶（AKP）阴性;油红O染色显示,支持细胞胞质内舍有大量的脂肪滴,核内有双极小体;丫啶橙染色证明支持细胞的细胞质中富含RNA,细胞核内的DNA含量也较高;罗丹明123染色证明支持细胞含有丰富的线粒体。结论：经两步酶消化法以及低渗处理后能得到纯度较高的鸡胚睾丸支持细胞;37℃、5％CO2的培养条件适宜支持细胞的生长;AKP染色、油红O、丫啶橙和罗丹明123等方法简单易行,适用于鸡胚睾丸支持细胞的鉴定。 相似文献
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采用玻璃化冷冻方法对野外采集的牦牛睾丸组织进行冷冻,通过HE(hematoxylin-eosin)染色分析发现玻璃化冷冻后的牦牛睾丸组织曲细精管结构保存较完好,曲细精管可见大量形态完好的各类生精细胞。采用台盼蓝染色检测细胞活率,发现冷冻复苏后细胞活率可达80.20%。分别通过生精细胞和精原干细胞标志蛋白DDX4和GFRA1免疫荧光染色发现复苏后培养14天后的生精细胞和精原干细胞的数量明显减少。通过RT-qPCR对复苏后不同实验处理组牦牛睾丸细胞标志基因的表达分析,发现培养30天的生精细胞中的精原干细胞标志基因Thy1和UCHL1的表达量显著增高。因此,玻璃化冷冻保存的牦牛睾丸组织中曲细精管及其生精细胞得到了较好的保护,该方法对于其他哺乳动物生精细胞的长久有效保存具有重要的参考价值。 相似文献
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以新生犊牛睾丸为实验对象,应用组合酶法进行支持细胞分离培养,并研究了冷冻保存后支持细胞的生长特性。结果表明:在细胞分离时,消化睾丸组织,分离曲细精管法所获得的细胞悬液中的有效细胞数高于组织剪碎法。支持细胞体外培养,4 h后开始贴壁,3~4 d铺满培养皿底壁,传代后细胞生长较快,2 d即可增殖一代。HE染色,胞质染色较淡,而细胞核染色较深,呈圆形或椭圆形位于细胞质中央或偏位,核仁明显。采用10%FBS+10%DMSO的DMEM液做冷冻液,对细胞进行冷冻保存时,支持细胞的复苏率在65%以上。解冻后的支持细胞体外培养,4h开始有细胞贴壁,24h后大部分细胞贴壁,3~4d铺满培养皿底壁。 相似文献
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支持细胞（ＳｅｒｔｏｌｉＣｅｌ）以多种途径影响精子发生。随着研究手段、方法的改进，支持细胞的诸多作用，如细胞联系、内分泌调控以及与间质细胞、生精细胞之间的交互作用等，已经或正在逐步被证实和揭示出来。这些功能的精细协调，为精子发生创造了适宜的微环境，保证了精子发生的有序进程。 相似文献
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Histologic and immunohistochemical characterization of a testicular mixed germ cell sex cord-stromal tumor and a leydig cell tumor in a dog 总被引：1，自引：0，他引：1
Mixed germ cell sex cord-stromal tumors (MGSCTs) of the testis are rare in dogs. We describe the histopathology and immunohistochemical characteristics of an MGSCT associated with a Leydig cell tumor in a cryptorchid testis. Histologically, MGSCT consisted of two nodules of seminiferous tubules lined by germ cells and Sertoli cells in variable proportions. Germ cells had variable size and nuclear features, with frequent giant cells. Germ cells were evenly mixed with Sertoli cells or located in the center of tubules. Markers that labeled mainly germ cells and few or no Sertoli or Leydig cells were calretinin, KIT, and PGP 9.5. E-cadherin, GATA-4, inhibin-alpha (INH-alpha), and neuron-specific enolase (NSE) were predominantly detected in Sertoli cells, whereas melan A was particularly expressed in Leydig cells and vimentin in all three cell types. OCT3/4 was not detected in any cell type. Although more cases of canine MGSCT need to be examined, our results suggest that an immunohistochemical panel of E-cadherin, GATA-4, INH-alpha, KIT, NSE, PGP 9.5, and melan A will help distinguish the three main cell types in canine testicular germ cell and sex cord-stromal tumors. 相似文献
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Xinpeng Yang Yue Feng Yang Li Dake Chen Xuanyan Xia Jialian Li Fenge Li 《Reproduction in domestic animals》2021,56(3):416-426
Sertoli cells are the only somatic cells in the seminiferous epithelium which directly contact with germ cells. Sertoli cells exhibit polarized alignment at the basal membrane of seminiferous tubules to maintain the microenvironment for growth and development of germ cells, and therefore play a crucial role in spermatogenesis. Androgens exert their action through androgen receptor (AR) and AR signalling in the testis is essential for maintenance of spermatogonial numbers, blood–testis barrier integrity, completion of meiosis, adhesion of spermatids and spermiation. In the present study, we demonstrated that AR gene could promote the proliferation of immature porcine Sertoli cells (ST cells) and the cell cycle procession, and accelerate the transition from G1 phase into S phase in ST cells. Meanwhile, miR-124a could affect the proliferation and cell cycle procession of ST cells by targeting 3′-UTR of AR gene. Furthermore, AR bound to the RNF4 via AR DNA-binding domain (DBD) and we verified that RNF4 was necessary for AR to regulate the growth of ST cells. Above all, this study suggests that AR regulates ST cell growth via binding to RNF4 and miR-124a, which may help us to further understand the function of AR in spermatogenesis. 相似文献
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D E Corrier H H Mollenhauer D E Clark M F Hare M H Elissalde 《Veterinary pathology》1985,22(6):610-616
Dietary cobalt (265 ppm Co) induced polycythemia and consistent degenerative and necrotic lesions in the seminiferous tubules of rats. Cyanosis and engorgement of testicular vasculature on day 35 and thereafter was followed on day 70 by degenerative and necrotic changes in the germinal epithelium and Sertoli cells. Spermatogonia, primary spermatocytes and round spermatids were markedly affected, while elongated spermatids, spermatozoa, and sertoli cells were more resistant. Damaged tubules, often present side by side with normal tubules, contained multinucleated giant cells composed of degenerated and necrotic spermatocytes and/or spermatids, sloughed germinal and Sertoli cells, and calcified necrotic debris. Necrotic tubules were frequently collapsed and devoid of epithelium except for occasional spermatogonia and surviving Sertoli cells. Lesions were not observed in the Leydig cells, cauda epididymis or seminal vesicles. 相似文献
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Oatley JM Tibary A de Avila DM Wheaton JE McLean DJ Reeves JJ 《Journal of animal science》2005,83(3):604-612
Spermatogonial stem cell transplantation is a technique that has potential in livestock to enhance genetic gain and generate transgenic offspring through the male germ line. A means for depletion of endogenous germ cells in a recipient's seminiferous tubules is necessary for this technology to be applied. The objectives of this study were to evaluate several methods for depletion of endogenous germ cells in the testes of adult rams and to evaluate ultrasound-guided injections into the rete testes as a means for infusing a suspension into the seminiferous tubules. Sixteen adult rams were randomly divided into 4 treatment groups (n = 4 per group). Treatments consisted of active immunization against LHRH (IMM), localized testicular irradiation (IR), LHRH immunization + irradiation (IMM+IR), and untreated control. Serial bleedings were conducted pretreatment and monthly after treatment for 4 mo, at which time all rams were castrated. Both IMM and IMM+IR rams received exogenous gonadotropin in the form of Perganol weekly for 8 wk before castration to bypass the immunization. All rams also received an ultrasound-guided injection of PBS containing 0.4% trypan blue into the rete testis of one testicle before castration. Rams receiving IMM and IMM+IR treatments had higher (P < 0.05) average percentages of seminiferous tubule cross sections with depleted germ cells compared with controls. Serum testosterone was decreased (P < 0.05) in IMM and IMM+IR rams 1 mo after treatment and throughout the remainder of the study compared with controls and IR rams, which were not different from each other. Serum inhibin concentration was unchanged in all rams following treatment indicating that Sertoli cell function was unaltered. A greater (P < 0.05) average percentage of the total testicular area could be filled with the trypan blue solution by rete testis injection in IMM and IMM+IR rams. These data demonstrate the depletion of endogenous germ cells in adult ram testes without alteration of Sertoli cell viability and function that have potential as methods for preparing recipient animals for germ cell transplantation. 相似文献
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Although several methods of testicular biopsy have been proposed previously, testicular fine needle aspiration (FNA) has proved to be the simplest, the most rapid, inexpensive, and overall the least invasive technique for obtaining testicular biopsies. Testicular FNA is indicated for fertility investigations in stallions with oligozoospermia or azoospermia. It is also used for differential diagnosis of testicular enlargement. After sedation, the stallion’s testis is punctured to obtain testicular parenchyma samples containing cells mainly from the seminiferous epithelium. The material obtained is used to perform smears which are analyzed for identification and quantification of germ cells and Sertoli cells. The results are based on the presence of the cell types found in the smears and the proportions of Sertoli cells per germ cells. In addition to being a very useful diagnostic tool, testicular FNA is also used for follow-up examinations, as it is minimally invasive. 相似文献
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Khalaj M Abbasi AR Nishimura R Akiyama K Tsuji T Noguchi J Okuda K Kunieda T 《The Journal of reproduction and development》2008,54(3):225-228
Repro22 is an N-ethyl-N-nitrosourea (ENU)-induced mutation in mice showing depletion of both male and female germ cells. In the present study, we investigated the male phenotypes of the mutant mouse at the adult stage. The repro22/repro22 homozygous mice showed reduced body weights as well as markedly reduced testis weights. Histological examination of the testes at 4 and 10 months of age showed no germ cells in the seminiferous tubules of the affected testis while a number of Sertoli cells were observed in the tubules. In addition to the germ cell depletion, the testes of the affected mouse contained expanded intertubular spaces that were filled by Leydig cell-like interstitial cells. These interstitial cells were confirmed to be Leydig cells by immunohistochmical staining using anti-3beta-HSD antibody. The estimated number of Leydig cells in the affected testes at 10 months of age increased approximately 2 fold compared with those of normal testes. Furthermore, the plasma testosterone levels of the affected mice at 10 months of age were significantly higher than those of the normal mice. These findings indicated that the repro22/repro22 mouse developed hyperplasia of Leydig cells that was presumably caused by the absence of germ cells in the seminiferous tubules. 相似文献