Isolation and cultivation of equine corneal keratocytes, fibroblasts and myofibroblasts.
Isolation and cultivation of equine corneal keratocytes, fibroblasts and myofibroblasts.
复制标题
DOI：
10.1111/j.1463-5224.2009.00755.x
复制
发表时间：
2010-01
影响因子：
1.6
通讯作者：
Mohan RR
中科院分区：
文献类型：
--
作者：
Buss DG;Giuliano EA;Sharma A;Mohan RR
目的：建立马角膜创面愈合的体外模型。为了实现这一目标，我们开发了一种分离和培养马角膜角质细胞、成纤维细胞和肌成纤维细胞的方法。马的角膜钮扣是无菌采集的，从健康的研究马进行人道安乐死的原因与本研究无关。在安乐死之前，由委员会认证的兽医眼科医生进行裂隙灯生物显微镜检查，以确保所有样本都是从没有前段疾病的马身上采集的。采用机械分离法分离马角膜基质，培养基质亚切片。采用定制培养基，在不同的培养条件下促进角膜基质细胞向角质细胞、成纤维细胞和肌成纤维细胞的生长和分化。应用细胞培养技术成功建立了马角膜角质细胞、成纤维细胞和肌成纤维细胞的分离培养方法。采用α -平滑肌和f -肌动蛋白免疫组化染色明确区分三种细胞类型。马角膜基质角质细胞，成纤维细胞和肌成纤维细胞可以预测地分离和体外培养使用该方案。
To establish an in vitro model for the investigation of equine corneal wound healing. To accomplish this goal, a protocol to isolate and culture equine corneal keratocytes, fibroblasts and myofibroblasts was developed. Equine corneal buttons were aseptically harvested from healthy research horses undergoing humane euthanasia for reasons unrelated to this study. Slit-lamp biomicroscopy was performed prior to euthanasia by a board-certified veterinary ophthalmologist to ensure that all samples were harvested from horses free of anterior segment disease. Equine corneal stroma was isolated using mechanical techniques and stromal sub-sections were then cultured. Customized media at different culture conditions was used to promote growth and differentiation of corneal stromal cells into keratocytes, fibroblasts and myofibroblasts. Cell culture techniques were successfully used to establish a method for the isolation and culture of equine corneal keratocytes, fibroblasts and myofibroblasts. Immunohistochemical staining for alpha-smooth muscle and F-actin was used to definitively differentiate the three cell types. Equine corneal stromal keratocytes, fibroblasts and myofibroblasts can be predictably isolated and cultured in vitro using this protocol.
登录
查看更多内容
影响因子：
4.4
作者：
Sharma A;Mehan MM;Sinha S;Cowden JW;Mohan RR
通讯作者：
Mohan RR
DOI：
10.1073/pnas.93.9.4219
发表时间：
1996-04-30
影响因子：
11.1
作者：
Masur, SK;Dewal, HS;Petridou, S
通讯作者：
Petridou, S
影响因子：
3.4
作者：
Mohan, RR;Hutcheon, AEK;Wilson, SE
通讯作者：
Wilson, SE
影响因子：
2.8
作者：
Leccisotti, Antonio
通讯作者：
Leccisotti, Antonio
影响因子：
1.6
作者：
Haber, M;Cao, ZY;Provost, PJ
通讯作者：
Provost, PJ