外阴组织是由正常的人源性阴道上皮细胞和树突状细胞培养而成. 其高度分化的结构与体内组织相似，是女性卫生毒性研究的理想选择, vaginal care, and microbicide products. Four types of EpiVaginal are offered:
1) VEC-100 (Figure 1): An epithelial tissue containing epithelial VEC cells,
2) VLC-100(图1):上皮组织，包含上皮VEC和具有免疫能力的树突状细胞,
3) VEC-100- ft(图2):VEC-100的全层版本，包括VEC上皮细胞和含有固有层的成纤维细胞, and
4) VLC-100-FT(图2):voc -100- ft的免疫能力版本，其中包括树突状细胞.
Figure 1: H&E染色A) VEC-100的组织学(福尔马林固定)横截面，体外重建含有正常人VEC细胞的上皮组织模型, and B) vaginal explant tissue. 体外和体内组织均显示有核的基底细胞层和上基底细胞层，随后是细胞核缺失和细胞充满糖原的细胞层.
图3:细胞角蛋白免疫染色的VEC-100组织的福尔马林固定横截面, CK13, CK14, & CK18. Basal cells are stained by CK14 and supra-basal cells by CK13; no cells are stained by CK18.
图4:暴露于含有常见杀精剂的配方18小时后，阴道外全层(VEC-100-FT)组织模型的活力(A)和细胞因子释放(B), nonoxynol-9 (N9): a) PBS control, b) KY jelly (KY), c) KY with 2% N9 (2% N9), and d) 0.1% N9 in PBS (0.1% N9). Note: As the tissue viability decreases, the IL-1β release increases.
图5:局部暴露于60(24小时)后VLC-100组织的HIV-1感染,000 CPM of HIV-1 ADA or NL4-3 virus. After exposure, tissues were washed 3X and then cultured for an additional 24 hr. 然后提取细胞DNA，使用HIV-1 gag特异性引物对检测HIV-1转录本.
图6:RT - PCR显示:A)器官型阴道-宫颈外(VEC)组织模型和B)人宫颈组织外植体中toll样受体(TLR)的表达.
杀菌剂测试，性病感染研究，女性卫生，炎症，药物输送
Irritation Testing
使用外阴组织测量暴露于女性卫生用品和其他产品后的组织活力和细胞因子释放.
EpiVaginal Irritation Protocol
EpiVaginal为女性产品安全测试以及HIV-1和其他性传播疾病的研究提供了一个体外研究工具. 其组织结构和细胞生理与体内阴道上皮组织密切相似, making it ideal for toxicology and efficacy testing, as well as a tool for biological research. Browse our reference library to see how researchers have used our EpiVaginal tissue in these areas of study.
Tissue
Kits: The EpiVaginal kit VEC-100 contains 24 tissues, VEC-112 contains 12 tissues and VEC-606 contains 6 tissues. (Tissue “kits” contain tissues, a small amount of culture medium, and plasticware; contact bbin体育官方网站 for specific kit contents.)
Substrate: Single well tissue culture plate inserts are used.
VEC-100: Nunc™ single well tissue culture plate inserts. Pore Size = 0.4 µm, Inner Diameter = 0.80 cm, Surface Area = 0.5 cm2.
VEC-606: Polycarbonate single well tissue culture plate inserts, Nunc™. Pore Size = 0.4 µm, Inner Diameter = 2.5 cm, Surface Area = 4.2 cm2.
Culture: At air liquid interface.
Histology: VEC-100: 10-16 cell layers of non-cornified tissue.
Lot Numbers: 每个技术员每周生产的纸巾批号被分配到一个特定的批号. Typically, there are multiple lot numbers for any given week’s tissue production. 在批号的末尾附加一个字母，以区分给定批号中的单个试剂盒. All tissue kits within a lot are identical with regard to cells, medium, handling, culture conditions, etc.
Shipment: At 4°C on medium-supplemented, agarose gels.
Shipment day: Monday.
Delivery: Tuesday morning via FedEx priority service (US). Outside US: Tuesday-Thursday depending on location.
Shelf life: 包括运输时间在内，纸巾在使用前可在4°C下保存4天. However, extended storage periods are not recommended unless absolutely necessary. In addition, 如果组织在同一天连续使用，将获得最佳的再现性, e.g. Wednesday morning following overnight storage at 4°C.
Length of experiments: 培养可持续至少1周，正常形态保持良好. Cultures must be fed every other day with 5.0 ml of maintenance medium (VEC-100-MM). 细胞培养插入放置在培养架(零件号MEL-STND)或垫圈(零件号EPI-WSHR)在6孔板允许使用5.0 ml.
Alternative tissues:
VEC-100- ft:除了在胶原/阴道成纤维细胞底物(固有层)上生长的组织外，与VEC-100相似。.
VEC-112-FT: Similar to VEC-100-FT except 12 tissues.
VLC-100: Similar to VEC-100 except tissues contain Langerhans cells.
VLC-112: Similar to VLC-100 except 12 tissues.
VLC-100- ft:除了在胶原/阴道成纤维细胞底物(固有层)上生长的组织外，与VLC-100相似.
VLC-112-FT: Similar to VLC-100-FT except 12 tissues.
Cells
Type: 正常人宫颈外周细胞(NHEC)分化为非角化组织, vaginal-ectocervical phenotype (part number VEC-100).
Genetic make-up: Single donor.
Derived from: adult human ectocervical tissue.
Screened for: HIV, Hepatitis-B, Hepatitis-C, mycoplasma, bacteria, yeast, fungi.
Medium
Base medium: Dulbecco’s Modified Eagle’s Medium (DMEM) and F12 Medium.
Growth factors/hormones: Epidermal growth factor and other proprietary factors.
Serum: None.
Antibiotics: Gentamicin 5 µg/ml (10% of normal gentamicin level).
Anti-fungal agent: Amphotericin B 0.25 µg/ml.
pH Indicator: Phenol red.
Other additives: Proprietary.
Alternatives: Phenol red-free, antibiotic-free, anti-fungal-free medium and tissue are available. Agents are removed 3 days prior to shipment.
Maintenance medium: VEC-100-MM
Quality Control and Sterility
Visual inspection: All tissues are visually inspected and if physical imperfections are noted, tissues are rejected for shipment.
End-use testing: Tissue viability of each lot is determined by an MTT assay. Tissues are exposed to 1.0% Triton X-100 for 30, 60, and 120 minutes. 使用MTT测定降低组织活力(ET-50)所需的暴露时间. The time of exposure needed to reduce the viability to 50% (ET-50) must be 0.92 – 1.67 hours. 应客户要求，将QC检测结果汇总提供给客户参考. (For TEER measurements, bbin体育官方网站 has successfully used the WPI Endohm System.)
Sterility: All media used throughout the production process is checked for sterility. 维持培养基加或不加抗生素孵育1周，检查无菌性. 用于运输的24孔板上的琼脂糖凝胶也孵育1周，并检查任何污染迹象.
Screening for pathogens: 所有细胞都经过筛选，使用PCR检测HIV、乙肝和丙肝呈阴性. 然而，没有任何已知的测试方法可以完全保证细胞是无病原体的. Thus, 这些产品和所有人类衍生产品应按照CDC-NIH手册中建议的BSL-2(生物安全等级2)或更高级别处理, “Biosafety in microbiological and biomedical laboratories,” 1998. 如需进一步帮助，请联系您的现场安全专员或bbin体育官方网站技术服务.
Notification of lot failure: If a tissue lot fails our QC or sterility testing, 我们会通知顾客，并在适当情况下免费更换纸巾. Because some of our QC and sterility testing is done post-shipment, notification will be made as soon as possible. (Under normal circumstances, ET-50 failures will be notified by Thursday, 5 p.m.; sterility failures will be notified within 8 days of shipment.)
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