成纤维细胞的

The invention discloses a connecting city white duck blastodisk fiber cell system with connecting city white duck embryo as material with preservation number at CGMCC No.1877 in cellular biology domain, which is characterized by the following: the fiber cell does not possess epithelial cell with high purity; the quality of freezing cell is stable; the active ratio can reach between 93.5% and 96.8%; the passage growth is stable and fit for big scale culture.

本发明利用连城白鸭胚胎作为材料，进行初代培养、传代培养及细胞冻存等研究。最终获得高活率、高纯度的连城白鸭胚成纤维细胞系，其保藏编号为CGMCC No.1877属于细胞生物学领域。本发明培养的成纤维细胞无上皮细胞等，细胞纯度高；冻存后细胞质量稳定，活率可达到并维持在93.5％～96.8％之间，传代生长稳定，适合大规模培养。

We identified that in vitro ASM promotes fibrocyte chemotaxis and chemokinesis (distance of migration after 4.5 hours, 31 mum [2.9 mum] vs 17 mum [2.4 mum], P =.0001), which was in part mediated by platelet-derived growth factor (mean inhibition by neutralizing antibody, 16% [95% CI, 2% to 32%], P =.03) but not by activation of chemokine receptors.

该研究首次证明：在哮喘中，成纤维细胞存在于哮喘患者气道平滑肌间隔，并可以促进纤维细胞迁移。成纤维细胞在哮喘患者气道平滑肌增生及气道功能障碍过程的所起的作用仍有待研究。

Results:Expression of elastase mRNA has been found in the endothelial cells,the medial smooth muscle cells and the adventitial fibroblasts of the abdominal aorta,the lymphocytes,monocytes in blood,the tracheal hyaline cartilaginous cells,the glandular cells of the pancreas,the epithelial cells of the parotid gland and submaxillary gland,the hepatoeytes,the endothelial cells of the liver sinusoid wall,the goblet cells of the mucous membrane of the small intestine,the cardiac myocytes,the renal interstitial fibroblasts,the alveolar epithelial cells,the cerebral glial cells,the fibroblasts of the dermis oorium of the skin,the primary spermaocytes,the secondary spermaocytes and sperm in the seminfferous tubule of the testis,the lymphocytes in the spleen and thymus.

结果正常大鼠腹主动脉的内皮细胞、中膜平滑肌细胞以及血管外膜成纤维细胞，血液细胞中的淋巴细胞、单核细胞，气管透明软骨细胞，胰腺的腺细胞、腮腺、颔下腺上皮细胞，肝细胞、肝窦壁的内皮细胞，小肠黏膜杯状细胞，心肌细胞，肾间质的纤维母细胞，肺泡上皮细胞，大脑胶质细胞，皮肤真皮纤维母细胞，睾丸曲精细管内的初级精母细胞、次级精母细胞以及精子，脾脏以及胸腺的淋巴细胞等，均有弹力蛋白酶mRNA的表达。

However, whether the human fibroblasts behave in a similar way has not been reported to our knowledge. In the present study fibroblasts collected from human foreskin were cultured in vitro so as to investigate into the fibro blastic osteogenetic role as well as its mechanism.

本实验包括四个部分：1、人皮肤成纤维细胞体外培养成骨的实验研究。2、猪骨形态形成蛋白（pBMP的提取及生物活性测定。3、pBMP对成纤维细胞成骨影响的实验研究。4、成纤维细胞体外培养过程中骨钙素变化。

Special staining methods, such as Masson and the Van Gieson staining were used to study the distribution of collogen fibers and elastic fibers. ResultsBy HE staining, the subepithelial connective tissues and vessels in the pterygium were more prominent than normal conjunctival tissues. An amorphous subepithelial superficial hyalinized zone and coarse eosinophilic granular materials were observed in the pterygia, but they were not found in normal conjunctival specimens. Coarse fibers were visible only in the deeper subepithelial connective tissues of pterygial samples. With Masson′s staining, the dense staining of collagen fibers was also more prominent in the pterygium than in the subepithelial connective tissues of normal conjunctiva. Abnormal collagen fibers were visible in the deeper sub-epithelial connective tissues of pterygial samples. With Van Gieson staining, abnormal collagen fibers were visible in the deeper subepithelial connective tissues. Dark coarse elastic fibers were found in the abnormal fibers only in the subepithelial deep connective tissues of pinguecula in the pterygia but not in the conjunctiva. With immunohistochemistry staining, MMP-3 was strong in the pterygial epithelium, moderate in fibroblast and absent from pterygial vascular walls. LN was strongly expressed in the blood vessel wall, moderately in the epithelial basement membrane and absent from the entire stroma.

结果HE染色：翼状胬肉组织上皮下基质中存在结缔组织的增生和血管形成；基质浅层存在一无定形物质透明区及粗糙的颗粒样嗜酸性物质，在翼状胬肉体部深层基质中存在粗糙的纤维组织；正常球结膜组织细胞排列整齐；基质为疏松结缔组织，胶原纤维平行排列，其间可见成纤维细胞，散在少量中性粒细胞、毛细血管；Masson染色：翼状胬肉浅层基质中存在致密的胶原纤维染色，深层基质中的胶原纤维存在变性样改变；VG染色：翼状胬肉组织深层基质中存在大量变性的胶原纤维，其间夹杂黑色的弹性纤维；免疫组化染色法：MMP-3在翼状胬肉上皮细胞中呈强表达，成纤维细胞中呈中等强度表达，血管内皮细胞中未见表达；LN在血管壁中呈强表达，在上皮细胞基底膜中呈中等强度表达，在整个基质中未见明显表达；col Ⅲ在整个翼状胬肉基质中呈强表达。

Methods Pterygial samples were extracted and collected and the pterygial endothelial cells and pterygial fibroblasts were cultured alone, conditional and co-cultured to form different culture systems. The methods included that to select the suitable intensity of ultraviolet by MIT, to detect the changes of curves of growth about two kinds of cells by MIT and to explore the developments of protein and RNA of vascular endothelial growth factor and fibroblast growth factor-basic in three culture systems under ultraviolet whose intensity is 20 mJ/cm^2 by ELISA and RT-PCR.

收集翼状胬肉标本，采用血管内皮细胞和成纤维细胞单独培养、条件培养和共同培养的方法构建体系，用MTT法选择紫外线照射细胞的适宜强度；采用MTT法绘制强度20mJ/平方公分紫外线照射下细胞生长曲线；采用ELISA和RT-PCR检测紫外线照射下3种体系中细胞上清液和细胞中血管内皮细胞生长因子和成纤维细胞生长因子的蛋白和RNA含量变化。

Results The results showed that fibroblasts were the main source of extracellular matrix production in bile duct wall. The phenotype of fibroblasts in inflammatory strictured bile duct wall changed obviously, quiescent fibroblasts were activated and transformed to myofibroblasts, with massive proliferation.

结果 成纤维细胞是胆管壁中合成胶原的主要细胞；炎性狭窄胆管壁的纤维化增厚明显，其成纤维细胞表型改变，由相对静止转化为功能活跃状态，并进一步向肌成纤维细胞转化，且大量增殖。

After γ ray treatment, two kinds of cells were incubated on 6-well gelatin-coated plate with density of 2.5×104/cm3. hESCs were cultured on HAFi or MEF feeder cells containing β-mercaptoethanol DMEM/F12 and basic fibroblast growth factor.

取人胚胎干细胞，分别接种在小鼠胚胎成纤维细胞或永生化人成纤维细胞饲养层上，加入含β-巯基乙醇的DMEM/F12培养基，使用前添加碱性成纤维细胞生长因子。

The hFCECs were cultured by sticking tissues piece method that the cornea were divided into the limbus and central tissue piece and digestive method,respectively.For digestive method,the digestive juice of 5mg/ml DispaseⅡ+0.25%trypsin was chosed.D/F12 with 10%fetal bovine serum and 100 U/ml penicillin and streptomycin were used in this study,the culture condition was 37℃and 5%CO2,and culture medium changed every three days2、Passage of hFCECsAfter more than 80%confluenced,cells from corneal limbus hFCECs were digested for 5-15min with different concentrations of trypsin/EDTA at 37℃,and then passaged at a ratio of 1:2.The cells were subcultured on empty plates,mouse 3T3 fibroblast feeder layer,fetal corneal stromal cell feeder l ayer or HTK feeder layer respectively.And D/F12, mouse 3T3 fibroblast conditioned medium,fetal corneal stromal cells conditioned medium or HTK conditioned medium with 10%FBS and 100 U/ml penicillin and streptomycin were used respectively as the culture medium.And the culture medium were changed every three days as well.

方法一、人胎儿角膜上皮原代和传代培养1、原代培养严格无菌操作获取人胎儿角膜片，采用组织块贴壁法、酶消化法原代培养人胎儿角膜上皮细胞。2、传代培养角膜缘部的细胞生长融合达80%以上，不同稀释浓度的胰酶/EDTA消化液消化传代分别接种于空板、小鼠3T3成纤维细胞饲养层、胎儿角膜基质细胞饲养层及HTK饲养层，培养液分别采用D/F12、小鼠3T3成纤维细胞条件培养液、胎儿角膜基质细胞条件培养液、HTK条件培养液。

In order to confirm this suspect, we choosed a cultured rat granulation tissue fibroblasts model in vitro and applied RT-PCR and cell-ELISA technology to detect the changes of firbroblasts intrinsic EGF, bFGF, TGFβ-1 with their receptors gene expressions and protein synthesises after stimulated by SP. Our purpose was to explore the possible effects and patterns imposed by SP on fibroblasts intrinsic growth factors and their receptors expressions, which maybe offer theoretical basis for promoting wound healing via improving nervous functions and regulating neuropeptides secretions.

为进一步验证这一推测，同时排除在体多因素干扰，我们采用了一种大鼠肉芽组织成纤维细胞体外培养模型，采用RT-PCR与细胞ELISA技术，检测SP刺激成纤维细胞后，成纤维细胞内源性EGF、bFGF、TGFβ-1及其受体基因表达和蛋白合成的改变情况，探讨SP对成纤维细胞内源性生长因子及其受体表达的影响以及方式，以期为经由改善神经功能、调节神经肽分泌途径促进伤口愈合提供理论依据。

For a big chunk of credit-card losses; the number of filings (and thus charge-off rates) would be rising again, whether

年美国个人破产法的一个改动使得破产登记急速下降，而后引起了信用卡大规模的亏损。

Eph. 4:23 And that you be renewed in the spirit of your mind

弗四23 而在你们心思的灵里得以更新