bbin体育官方网站的黑色素皮肤系统由正常的, human-derived epidermal keratinocytes (NHEK) and melanocytes (NHM) cultured to form a multilayered, 人表皮的高度分化模型. 共培养中的NHM发生黑色素形成，导致组织色素沉着. MelanoDerm is produced using serum free medium without artificial stimulators of melanogenesis such as TPA and IBMX. 培养物生长在空气-液体界面的组织培养插入物上, 允许局部应用皮肤美白剂或自晒黑剂. MelanoDerm provides a useful in vitro means to evaluate cosmetic and pharmaceutical agents designed to modulate skin pigmentation.
MelanoDerm exhibits in vivo-like morphological and ultrastructural characteristics which are uniform and highly reproducible. NHM localized in the basal cell layer of MelanoDerm are dendritic and spontaneously produce melanin granules which progressively populate the layers of tissue. 培养3周, MelanoDerm tissues become increasingly pigmented with retention of normal epithelial morphology. Tissues containing NHM derived from black donors show increased pigmentation versus those containing Caucasian-derived or Asian-derived NHM; all three types of cultures are distinctly darker than NHM-free tissues (EpiDermTM). The topical application of known inhibitors of melanogenesis significantly reduce melanin production and macroscopic darkening. Conversely, 组织内的NHM会对已知的黑色素生成刺激物产生反应, 如α-促黑素细胞激素和β-成纤维细胞生长因子, 产生的组织比未经处理的组织变黑得更快. MelanoDerm’s straightforward assays has been incorporated as an integral testing strategy in laboratories for nearly 20 years. 有几十个技术参考资料可用, this growing body of data demonstrates that MelanoDerm provides an inexpensive and effective means of assessing skin pigmentation while avoiding species extrapolation and the use of laboratory animals.
MelanoDerm provides an inexpensive and effective means of assessing skin pigmentation while avoiding species extrapolation and the use of laboratory animals. Simple protocols and the evaluation of early cellular endpoints allow scientists to acquire data in days, 不是几周或几个月.
美白
A quantitative assay to determine melanin content in the tissue is performed using the SOLVABLE™ melanin assay. 该试验可用于测试最终化妆品配方的皮肤美白功效.
色素的研究
MelanoDerm已被用于研究皮肤色素沉着的生理机制.
Tissue:
Kit: Standard 黑种人™ kit (MEL-300) consists of 24 individual tissues, each tissue 9 mm in diameter. (纸巾“包”包含纸巾, 少量培养基, and plasticware; contact bbin体育官方网站 for specific kit contents.)
Substrate: Chemically modified, collagen-coated, 9 mm ID single well tissue culture plate inserts are used (e.g. Millicell CM或Nunc聚碳酸酯单孔组织培养板插入件).
培养:在气液界面.
播种率: Nhm: nhek = 1/10. 1/30的比例可根据要求提供.
Histology: 8-12个细胞层(基底层、棘层和颗粒层).
角质层: 10-15层(基于TEM).
很多数字: 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. 通常，任何一周的纸巾生产都有多个批号. A letter of the alphabet is appended to the end of the lot number to differentiate between individual kits within a given lot of tissues. 所有组织试剂盒在细胞方面都是相同的, medium, handling, 培养条件, etc.
Shipment: 在4°C培养基中，琼脂糖凝胶在24孔板中.
装运: 每星期一. 如有特殊要求，也可在星期四装运.
Delivery: 周二上午通过联邦快递优先服务(美国). 美国以外:周二至周三，视地点而定.
保质期: 包括运输时间在内，纸巾在使用前可在4°C下保存6天. 但是，除非必要，否则不建议延长储存期限. 除了, 如果组织在同一天连续使用，将获得最佳的再现性, e.g. 周二下午或之后在4°C下过夜(周三上午).
实验时长: Cultures can be continued for at least 2 weeks with good retention of normal epidermal morphology. 培养物必须每隔一天用5.0毫升新维护介质(NMM)或NMM-113. Cell culture inserts are placed atop culture stands (MEL-STND; see photo below) or washers (EPI-WSHR) in 6-well plates to allow use of 5.0 ml.
替代组织:
梅尔- 300: 与MEL-300相同，除了只有1个组织，而不是24个组织.
梅尔- 300 b - frzn ea: 经过3次冻融循环的MEL-300-B组织. 设计用于黑色素测定的对照.
MEL-301: 比MEL-300少培养3天，比MEL-300分化少
Mel-312 (-a， -b， -c): 与MEL-300 (-A， -B， -C)相同，除了每个试剂盒12个组织而不是24个.
Mel-606 (-a， -b， -c): 与MEL-300 (a)相同, -B, -C)除了每包6张纸巾, 每个组织在直径较大的细胞培养插入物(ID = 22 mm)中培养.
mel -300英尺(-a， -b， -c): Full-thickness human skin tissue model composed of fibroblast-containing collagen matrix (dermis); normal human epidermal keratinocytes and normal human melanocytes (epidermis). 每包24张纸巾.
Cells:
Types: a) Normal human epidermal keratinocytes (NHEK); b) Normal human melanocytes (NHM).
基因组成: NHEK和NHM分别来自不同的捐赠者. 含有匹配的NHEK和NHM的组织可根据特殊要求提供.
民族起源: 含NHM的白种人黑色素皮(MEL-300-C), 黑色(梅尔- 300 b), 以及亚洲(MEL-300-A)捐助者.
来自: Neonatal-foreskin组织.
选择: 有限数量的NHEK菌株 & NHM来自成人乳房组织.
筛选: 艾滋病毒，乙型肝炎和丙型肝炎.
Medium:
基础培养基: Dulbecco 's Modified Eagle 's Medium (DMEM).
生长因子/激素: 表皮生长因子, insulin, 氢化可的松, 以及其他专有的表皮分化刺激物.
Serum: None.
抗生素: 庆大霉素5µg/ml(为庆大霉素正常水平的10%).
抗真菌剂: 两性霉素B.25 µg/ml.
酸碱指示剂: 酚红.
其他添加剂: 用于增强表皮屏障形成的脂质前体(专有).
选择: 无酚红(MEL-300-B-PRF), 不需抗生素(梅尔- 300 b -沛富), anti-fungal-free(梅尔- 300 b -等于off), 不含氢化可的松的培养基和组织(MEL-300-B-HCF)可用. 代理商至少在发货前3天被移除.
维护中: 有四种维护介质:
对于长期的实验，其中色素沉着和组织学是重要的, EPI-100-NMM的使用, epi - 100 nmm - 113, 或EPI-100-LLMM.
EPI-100-NMM(新维持培养基)含有保存组织学的KGF, 但不含任何直接促进色素沉着的因素.
epi - 100 nmm - 113含有KGF, β-FGF, 和α-MSH，促进色素沉着和组织保持.
长寿命维护介质EPI-100-LLMM含有α-MSH和β-FGF，但不含KGF. Tissues will darken most rapidly in LLMM, less rapidly in NMM-113, and least rapidly in NMM.
当使用NMM或NMM-113时，可以获得最佳的组织学保留, 培养物被喂饱了.每48小时0毫升培养基.
第四种媒介, epi - 100毫米, 是否为基础培养基，可用于短期毒理学试验. 它不含任何促进色素沉着的因素. epi - 100毫米 should not be used for experiments in which tissue histology is an important endpoint.
质量控制和无菌:
目视检查: 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. Select tissues are examined microscopically to insure that melanocytes are visible (for MEL-300-B and MEL-300-A tissues) and that they are evenly distributed throughout the tissue.
最终用途测试: Tissues are continued in EPI-100-LLMM to insure that macroscopic darkening of tissues occurs. 将组织暴露在1% Triton X-100中5小时. The viability at 5 hrs must exceed 45% as determined by the MTT assay (See bbin体育官方网站 EpiDerm™ ET-50 protocol) for each lot of tissue.
不育: 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.
病原体筛查: 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.
不合格批件通知: 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QC和无菌检测是在出货后完成的, 在一般情况下，我们会尽快发出通知, 可行性失败将在周三下午5点前通知.m.; sterility failures will be notified within 8 days of shipment).
新订单
To ensure that the most appropriate products and accessories for your application are included with your initial order, bbin体育官方网站 strongly recommends a complimentary consultation with one of our highly trained Technical Specialists. 请联系乔纳森Oldach订购MelanoDerm产品.
乔纳森Oldach
bbin体育官方网站公司
joldach@gntextile.com
T:(508) 881-6771分机. 210
F: (508) 879-1532
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