成纤维细胞

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。

The notable proliferation was not observed by eyes in the local of injection. The infiltration of inflammation cells and mild proliferation of fibrocyte around dura mater was observed by HE stained in 4 and 8 weeks after injection. Infiltration and exudation of inflammation cells was observed by HE stained in epidural nerve root. Compared with group A, no changes of group B, C and D were observed under specific stained. Proliferation of type Ⅱ collagen fibers around dura mater was seen under immunohistochemical stained in 4 and 8 weeks after injection. There is no significant demyelination changes under LFB stained. The thickness and shape of the myelin sheath in epidural nerve root was not regular under transmission electronic microscopy in 4 and 8 weeks after injection. Fibroblast was also seen there. In nerve endometrium, macrophage could be seen under TEM, myelinated nerve fiber changed significantly, but nonmyelinated nerve fiber changed mildly. When 8 weeks, the changes of group D is smaller than the group B and C.

给药局部肉眼观察未见明显的纤维组织增生；HE染色可见B、C、D三组给药后四周及八周时硬膜内外均有炎细胞浸润，纤维细胞轻度增生，硬膜外神经根内有炎细胞浸润及炎性渗出；特殊染色B、C、D三组同A组相比未见有脊髓及神经根的改变；免疫组化染色，给药后四周及八周时，硬膜内外均有Ⅱ型胶原纤维增生；固兰染色B、C、D三组未见有明显脱髓鞘改变，与A组相比无明显异常改变；电镜观察B、C、D三组在给药后的四周及八周时，表现为硬膜外神经根内髓鞘厚薄不一，形状不规则，可见成纤维细胞，神经内膜中可见有巨噬细胞；粗大的有髓神经纤维变化明显，无髓神经纤维受累较轻；八周时电镜下D组改变较B、C两组为轻。

The cells were passaged no more than 5 generations,and the cell freezing density is about 3×106 cell/ml, sample numbers of each kind are between 30 and 60, conserved numbers are 150-180 ampules,biological characters of fibroblast were analysed according to the request of constructing cell line. The results were as follows: 1 Cultured cells'morpha is typical fibroblast,which is fusiformate or unregular triangle, cells is radiant,flaming or rotative during growing .Qingyuan flesh chicken's cytoplasmic tubercle is much more and shorter in contrast with Zang chicken and Langshan chicken,that can be regarded as different character from others.

细胞冻存代数在5 代以内，冻存密度为3×106个细胞/ml，每个品种的样本含量在30～60 个之间，保存管数达150～180 支，根据建系要求，对成纤维细胞的生物学性状进行分析，所得结果如下： 1 培养细胞形态为典型的成纤维状，为梭形或不规则三角形，细胞在生长时呈放射状、火焰状或旋涡状走形，清远麻鸡的细胞形态与藏鸡和狼山鸡相比，胞质向外伸出的突起较多，且较短。

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。

Rat dermal fibrobalasts and human neofetus dermal fibroblasts were cultured in vitro to evaluate the biocompatibility of the chitosan films.

为考察壳聚糖对皮肤成纤维细胞的相容性，采用流延法制备了纯壳聚糖膜和含甘油的壳聚糖膜，并在所制备的壳聚糖膜上进行了大鼠皮肤成纤维细胞和人胎儿皮肤成纤维细胞的体外培养。

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。

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。

Primary culture of goat mammary gland cells could be derived by outgrowth of migrating cells from fragment of tissue. Fibroblast and epithelial cells could be pured according to their different sensibility to trypsin. Complex cultures treated by 0.25% trypsin in Hanks' for 5~7min at 37℃, the dispersed cells were mainly fibroblasts,then the rest treated by 0.15% trypsin-0.02% EDTA in Hanks'for 5~8min at 37℃, the majority cells harvested were mammary epithelial cells.

用组织块培养法可获得良好的山羊乳腺细胞原代培养物；培养的山羊乳腺成纤维细胞比上皮细胞对胰蛋白酶更敏感，据此可在传代过程中将二者分离纯化，获得纯细胞系；混合培养物先用0.25%胰蛋白酶在37℃消化5~7min 所收集的细胞主要为成纤维细胞；再加0.15%胰蛋白酶-0.02鞹A 消化液在37℃继续消化5~6min 所回收的细胞绝大多数为上皮细胞，经过2~3 代，即可得到纯化的乳腺上皮细胞系。

Primary culture of goat mammary gland cells could be derived by outgrowth of migrating cells from fragment of tissue. Fibroblast and epithelial cells could be pured according to their different sensibility to trypsin. Complex cultures treated by 0.25% trypsin in Hanks' for 5~7min at 37℃, the dispersed cells were mainly fibroblasts,then the rest treated by 0.15% trypsin-0.02% EDTA in Hanks'for 5~8min at 37℃, the majority cells harvested were mammary epithelial cells.

用组织块培养法可获得良好的山羊乳腺细胞原代培养物；培养的山羊乳腺成纤维细胞比上皮细胞对胰蛋白酶更敏感，据此可在传代过程中将二者分离纯化，获得纯细胞系；混合培养物先用0.25%胰蛋白酶在37℃消化5~7min 所收集的细胞主要为成纤维细胞；再加0.15%胰蛋白酶-0.02%EDTA 消化液在37℃继续消化5~6min 所回收的细胞绝大多数为上皮细胞，经过2~3 代，即可得到纯化的乳腺上皮细胞系。

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。

Singer Leona Lewis and former Led Zeppelin guitarist Jimmy Page emerged as the bus transformed into a grass-covered carnival float, and the pair combined for a rendition of "Whole Lotta Love".

歌手leona刘易斯和前率领的飞艇的吉他手吉米页出现巴士转化为基层所涵盖的嘉年华花车，和一双合并为一移交&整个lotta爱&。

This is Kate, and that's Erin.

这是凯特，那个是爱朗。