| Title | Ex vivo 2D and 3D HSV-2 infection model using human normal vaginal epithelial cells |
| Authors | Zhu, Yaqi Yang, Yan Guo, Juanjuan Dai, Ying Ye, Lina Qiu, Jianbin Zeng, Zhihong Wu, Xiaoting Xing, Yanmei Long, Xiang Wu, Xufeng Ye, Lin Wang, Shubin Li, Hui |
| Affiliation | Wuhan Univ, Sch Basic Med Sci, Inst Med Virol, State Key Lab Virol, Wuhan 430071, Hubei, Peoples R China. Wuhan Univ, Shenzhen Inst, Shenzhen R&D Ctr, State Key Lab Virol, Shenzhen 518057, Guangdong, Peoples R China. Xiangyang 1 Peoples Hosp, Xiangyang 441000, Hubei, Peoples R China. Peking Univ, Shenzhen Hosp, Shenzhen 518036, Guangdong, Peoples R China. Hubei Maternal & Child Hlth Hosp, Wuhan 430070, Hubei, Peoples R China. Shenzhen Eye Hosp, Shenzhen 518040, Guangdong, Peoples R China. Wuhan Univ, Sch Basic Med Sci, Inst Med Virol, State Key Lab Virol, Wuhan 430071, Hubei, Peoples R China. Li, H (reprint author), Wuhan Univ, Shenzhen Inst, Shenzhen R&D Ctr, State Key Lab Virol, Shenzhen 518057, Guangdong, Peoples R China. |
| Keywords | three dimension (3D) ex vivo human normal vaginal epithelial cells (HNVEC) air-liquid interface (ALI) culture HSV-2 infection model HERPES-SIMPLEX-VIRUS MALIGNANT BREAST HTERT PROMOTER EXPRESSION TYPE-2 GROWTH IMMORTALIZATION RESPONSES ADHESION VACCINE |
| Issue Date | 2017 |
| Publisher | ONCOTARGET |
| Citation | ONCOTARGET.2017,8(9),15267-15282. |
| Abstract | Herpes simplex virus type 2 (HSV-2) infects human genital mucosa and establishes life-long latent infection. It is unmet need to establish a human cell-based microphysiological system for virus biology and anti-viral drug discovery. One of barriers is lacking of culture system of normal epithelial cells in vitro over decades. In this study, we established human normal vaginal epithelial cell (HNVEC) culture using co-culture system. HNVEC cells were then propagated rapidly and stably in a defined culture condition. HNVEC cells exhibited a normal diploid karyotype and formed the well-defined and polarized spheres in matrigel three-dimension (3D) culture, while malignant cells (HeLa) formed disorganized and nonpolar solid spheres. HNVEC cells had a normal cellular response to DNA damage and had no transforming property using soft agar assays. HNVEC expressed epithelial marker cytokeratin 14 (CK14) and p63, but not cytokeratin 18 (CK18). Next, we reconstructed HNVEC-derived 3D vaginal epithelium using air-liquid interface (ALI) culture. This 3D vaginal epithelium has the basal and apical layers with expression of epithelial markers as its originated human vaginal tissue. Finally, we established an HSV-2 infection model based on the reconstructed 3D vaginal epithelium. After inoculation of HSV-2 (G strain) at apical layer of the reconstructed 3D vaginal epithelium, we observed obvious pathological effects gradually spreading from the apical layer to basal layer with expression of a viral protein. Thus, we established an ex vivo 2D and 3D HSV-2 infection model that can be used for HSV-2 virology and anti-viral drug discovery. |
| URI | http://hdl.handle.net/20.500.11897/475023 |
| ISSN | 1949-2553 |
| DOI | 10.18632/oncotarget.14840 |
| Indexed | SCI(E) |
| Appears in Collections: | 深圳医院 |
