bbin体育官方网站’s EpiOral and EpiGingival tissues consist of normal, human-derived oral epithelial cells. 细胞已培养成多层, highly differentiated models of the human buccal (EpiOral) and gingival (EpiGingival) phenotypes. The tissues are cultured on specially prepared cell culture inserts using serum free medium and attain levels of differentiation on the cutting edge of in vitro cell culture technology. The EpiOral and EpiGingival tissue models exhibit in vivo-like morphological and growth characteristics which are uniform and highly reproducible.
Both EpiOral and EpiGingival express cytokeratin K13 in all layers except the most apical ones and weakly express cytokeratin K14 in the upper layers of the tissue (Figure 1). The tissues also produce the naturally occurring antimicrobial peptides called human beta defensins (HBDs). EpiOral组成性表达HBD-1和HBD-3，但不表达HBD-2(图2)。. EpiGingival weakly expresses HBD-3 in all layers except the uppermost ones and HBD-1 in the apical layers but does not express HBD-2 (Figure 3).
Lipid analysis of the explant tissues along with EpiGingival (GIN-100) and EpiOral (ORL-200) is shown below in Table 1. 口腔和口腔外植体只含有神经酰胺2(神经酰胺NS)。. 表面牙龈和牙龈组织含有神经酰胺1, 2, 和3(也称为神经酰胺EOS), NS, and EOHP/NP, respectively).
A growing body of data indicates that EpiOral and EpiGingival effectively provide an inexpensive, 非动物方法评估口腔刺激, toxicology, and pathology-related issues. 十二烷基硫酸钠(SDS)作用的代表性剂量-反应曲线, 牙膏中常用的表面活性剂, is shown in Figure 4.
Figure 4: Effect of SDS solutions (40L) on EpiOral (ORL-200) tissue viability following exposure for 1 hour. SDS的浓度选择在牙膏中通常存在的范围内(0.0 – 3.0%).
表1:体内和体外组织脂质分析. 体内的口腔和牙龈值是14个组织样本的平均值. 体外组织值来自单个批次.
粘膜和牙龈刺激
Measure oral or gingival epithelial tissue viability and cytokine release using a straightforward MTT assay or commercially available ELISA kit, 几天内就能得到确切的结果.
Drug Delivery
EpiOral and EpiGingival are useful in determining the absorption potential of final formulations in vitro.
Model for Oral Pathologies
EpiOral已被用于研究口腔念珠菌病, oral mucositis, 以及口腔黏膜的抗菌特性.
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Tissue:
Kits: EpiOral (ORL-200)和EpiGingival (GIN-100)试剂盒由24个组织组成. ORL-606和GIN-606试剂盒由6张纸巾组成. Tissue kits contain tissues, culture medium, 使组织平衡并培养24小时, and plasticware. 有关特定试剂盒内容，请联系bbin体育官方网站.
Substrate: ORL-200 and GIN-100: Collagen coated, 9 mm ID single well tissue culture inserts are used. Pore size = 0.4 µm, Inner Diameter = 0.875 cm. Surface area = 0.6 cm2. ORL-606和GIN-606:单孔组织培养插入物. Pore size = 0.4 µm, Inner Diameter = 2.5 cm. Surface area = 4.2 cm2.
Culture: At air-liquid interface.
Histology: ORL-200: 8-11个非角膜组织细胞层. GIN-100: 8-11层角膜组织.
Lot Numbers: 每周生产的纸巾批次被分配一个特定的批号. 所有组织试剂盒在细胞方面都是相同的, medium, handling, culture conditions, etc.
Shipment: 在4°C培养基中添加琼脂糖凝胶.
Shipment day: Monday.
Delivery: 周二上午通过联邦快递优先服务(美国). 美国以外:周二至周四视地点而定.
Shelf life: Including time in transit, tissues may be stored at 4°C for up to 96 hours prior to use. 但是，除非绝对必要，否则不建议延长储存期. In addition, the best reproducibility will be obtained if tissues are used consistently on the same day, e.g. 周三早上，在4°C的温度下过夜.
Length of experiments: 培养可持续至少1周，正常形态保持良好. 培养物必须每隔一天用5.0毫升维持培养基(ORL-200-MM或GIN-100-MM). Cell culture inserts are placed atop culture stands (MEL-STND) or washers (EPI-WSHR) in 6-well plates to allow the use of 5.0 ml.
Alternative tissues:
ORL-300-FT: Full thickness ORL-200. Oral epithelial tissue cultured on top of a collagen matrix containing gingival fibroblasts.
ORL-300-FT-LC: ORL-300-FT含朗格汉斯细胞.
GIN-300-FT: Full thickness GIN-100. Gingival epithelial tissue cultured on top of a collagen matrix containing gingival fibroblasts.
Cells:
EpiOral
Type: Normal human oral keratinocytes (NHOK) are differentiated into tissues with a non-cornified, buccal phenotype, part number ORL-200.
Genetic make-up: Single donor.
Derived from: Non-diseased, adult human oral tissue obtained from patients undergoing tooth extractions or from cadavers. Tissues are produced using cells derived from non-smokers who have no history of drug use or oral pathology.
Screened for: 艾滋病毒、乙型肝炎、丙型肝炎、支原体、细菌、酵母菌、真菌.EpiGingival
Alternate donors available. EpiOral produced using cells derived from patients with a history of smoking are available (ORL-200-D2).
EpiGingival
Type: Normal human oral keratinocytes (NHOK) are differentiated into tissues with a cornified, gingival phenotype, part number GIN-100.
Genetic make-up: Single donor.
Derived from:
Non-diseased, adult human oral tissue obtained from patients undergoing tooth extractions or from cadavers. Standard tissues are produced from cells derived from non-smokers who have no history of drug use or oral pathology.
Alternate donors available. EpiGingival produced using cells derived from patients with a history of smoking are available (GIN-100-D2).
Screened for: 艾滋病毒、乙型肝炎、丙型肝炎、支原体、细菌、酵母菌、真菌.
Medium:
EpiOral &EpiGingival
Base medium: Dulbecco 's Modified Eagle 's Medium (DMEM)
Growth factors/hormones: 表皮生长因子和其他专有因子
Serum: None
Antibiotics: Gentamicin 5 µg/ml
Anti-fungal agent: Amphotericin B 0.25 µg/ml
pH Indicator: Phenol red
Other additives: Proprietary
Alternatives: 不含酚红、不含抗生素、不含真菌的培养基和组织. 代理商在发货前3天撤离
Maintenance medium: ORL-200-MM or GIN-100-MM
质量控制和无菌:
Visual inspection: 所有组织都经过目视检查，如果发现物理缺陷, 纸巾被拒绝装运.
End-use testing:. 组织也暴露于1.0% Triton X-100 for various times and the exposure time required to reduce the tissue viability to 50% (ET-50) is determined using the MTT assay. QC规格:ORL-200: 34.85 < ET-50 < 105.78 min GIN-100: 5.47 hr < ET-50 < 10.42 hr.
Sterility: 在整个生产过程中使用的所有介质都要进行无菌检查. Maintenance medium is incubated with and without antibiotics for 1 week and checked for sterility. The agarose gel from the 24-well plate used for shipping is also incubated for 1 week and checked for any sign of contamination.
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. However, no known test method can offer complete assurance that the cells are pathogen free. Thus, these products and all human derived products should be handled at BSL-2 levels (biosafety level 2) or higher as recommended in the CDC-NIH manual, 微生物和生物医学实验室的生物安全,” 1998. For further assistance, please contact your site Safety Officer or bbin体育官方网站 technical service.
Notification of lot failure: 如果纸巾没有通过我们的质量检验或无菌检验, the customer will be notified and the tissues will be replaced without charge when appropriate. 因为我们的一些QC和无菌检测是在出货后完成的, 我们会尽快发出通知. 在正常情况下，ET-50故障将在周四下午5点前通知.m.; sterility failures will be notified within 8 days of shipment.
