bbin体育官方网站的黑色素皮肤系统由正常的, 人源性表皮角质形成细胞(NHEK)和黑色素细胞(NHM)培养形成多层, highly differentiated model of the human epidermis. The NHM within co-cultures undergo melanogenesis leading to tissue pigmentation. 黑素真皮是使用无血清培养基生产的，不含人工刺激黑素形成的物质，如TPA和IBMX. The cultures are grown on tissue culture inserts at the air-liquid interface, allowing for topical application of skin lighteners or self-tanning agents. 黑种人提供了一种有用的体外手段来评估化妆品和药物制剂，旨在调节皮肤色素沉着.
黑皮具有与活体相似的形态和超微结构特征，具有均匀性和高度可复制性. 位于黑色素真皮基底细胞层的NHM是树突状的，自发产生黑色素颗粒，逐渐填充组织层. 培养3周, 黑色素真皮组织变得越来越色素沉着，保持正常上皮形态. 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 (表皮TM). 局部应用已知的黑色素生成抑制剂可显著减少黑色素的产生和宏观变黑. 相反, NHM within the tissue will respond to known stimulants of melanogenesis, such as α-melanocyte stimulating hormone and β-fibroblast growth factor, to produce tissues which darken faster than untreated controls. 近20年来，黑种人的直接检测方法已被纳入实验室的整体检测策略. 有几十个技术参考资料可用, 越来越多的数据表明，黑种人提供了一种廉价而有效的评估皮肤色素沉着的方法，同时避免了物种外推和使用实验动物.
黑种人提供了一种廉价而有效的评估皮肤色素沉着的方法，同时避免了物种外推和使用实验动物. 简单的协议和早期细胞端点的评估使科学家能够在几天内获得数据, 不是几周或几个月.
皮肤亮
使用SOLVABLE™黑色素测定法进行定量测定以确定组织中的黑色素含量. 这个试验可以用来测试皮肤美白功效的最终化妆品配方.
色素的研究
黑种人 has been used to study the physiological mechanisms of skin pigmentation.
Search our library of technical references for more information.
组织:
工具: 标准黑种人™试剂盒(MEL-300)由24个单独的组织组成，每个组织直径9毫米. (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.)
基质: 化学修饰，胶原涂层，9毫米ID单孔组织培养板插入使用.
培养:在气液界面.
播种率: Nhm: nhek = 1/10. 可根据要求提供1/30的比例.
组织学: 8-12 cell layers (basal, spinous, and granular layers).
角质层: 10-15层(基于TEM).
很多数字: Tissue lots produced by each technician for each week are assigned a specific lot number. 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 in regards to cells, medium, 处理, 培养条件, 等.
装运: At 4°C on medium-supplemented, agarose gels in a 24-well plate.
装运: 每星期一. Shipment on Thursday is also available upon special request.
交付: Tuesday morning via FedEx priority service (US). Outside the US: Tuesday-Wednesday depending on location.
保质期: Including time in transit, tissues may be stored at 4°C for up to 6 days prior to use. However, extended storage periods are not recommended unless necessary. 除了, the best reproducibility will be obtained if tissues are used consistently on the same day, e.g. Tuesday afternoon or following overnight storage at 4°C (Wednesday morning).
实验时长: 培养可持续至少2周，保持良好的正常表皮形态. 培养物必须每隔一天用5.0 mL of either New Maintenance Medium (EPI-100-NMM-3) or epi - 100 nmm - 113. 细胞培养插入放置挂顶板(HNG-TOP-12，提供)，允许使用5.0 mL.
替代组织:
MEL-300-A(或-B或-C)-EACH: Same as MEL-300 except only 1 tissue, not 24 tissues.
MEL-300-A(或-B或-C) -FRZN-EA: MEL-300-B tissue that has been subjected to 3 freeze/thaw cycles. Designed to serve as a control for melanin assay.
Mel-301 (-a， -b， -c): Cultured 3 days less than MEL-300, less differentiated than MEL-300
Mel-312 (-a， -b， -c): Identical to MEL-300 (-A, -B, -C) except 12 tissues per kit instead of 24.
Mel-606 (-a， -b， -c): 与MEL-300 (a)相同, -B, -C)除了每包6张纸巾, each tissue cultured in a larger diameter cell culture insert (ID = 22 mm).
mel -300英尺(-a， -b， -c): Full-thickness human skin tissue model composed of fibroblast-containing collagen matrix (dermis); normal human epidermal keratinocytes, 正常的人类黑素细胞(表皮). 每包24张纸巾.
细胞:
类型: a) Normal human epidermal keratinocytes (NHEK); b) Normal human melanocytes (NHM).
基因组成: NHEK and NHM are from individual, but different donors. Tissue containing matched NHEK and NHM are available upon special request.
民族起源: 黑种人 containing NHM from Caucasian (MEL-300-C), 非裔美国人(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.
介质:
基础培养基: Dulbecco 's Modified Eagle 's Medium (DMEM).
生长因子/激素: 表皮生长因子, 胰岛素, 氢化可的松, and other proprietary stimulators of epidermal differentiation.
血清: 没有一个.
抗生素: Gentamicin 5 µg/mL (10% of normal gentamicin level).
抗真菌剂: 两性霉素B.25µg / mL.
酸碱指示剂: 酚红.
其他添加剂: Lipid precursors used to enhance epidermal barrier formation (proprietary).
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), and 氢化可的松-free medium and tissue (MEL-300-B-HCF) are available. Agents are removed at least 3 days prior to shipment.
维护中: 建议使用三种维护介质.
For long-term experiments in which pigmentation and histology are important, EPI-100-NMM-3的使用, epi - 100 nmm - 113, 或epi - 100 - llmm - 3所示.
EPI-100-NMM-3(新维持培养基)含有保留组织学的KGF，但不含任何直接促进色素沉着的因素.
epi - 100 nmm - 113含有KGF, βfgf, and α-MSH and promotes pigmentation as well as retention of histology.
长寿命保养介质, epi - 100 - llmm - 3所示, 含有高水平的α-MSH和βfgf, and also contains KGF at the same level as in EPI-100-NMM-3 and EPI-100-NNM-113. LLMM-3中组织变暗最快，NMM-113变暗较慢，NMM-3变暗最慢.
The best retention of histology will be obtained when the cultures are fed with 5.每48小时0 mL培养基.
质量控制和无菌:
目视检查: All tissues are visually inspected and if physical imperfections are noted, 纸巾被拒绝装运. 在显微镜下检查选择的组织，以确保黑色素细胞可见(对于MEL-300-B和MEL-300-A组织)，并确保它们均匀分布在整个组织中.
最终用途测试: Tissues are continued in culture to ensure that macroscopic darkening of tissues occurs. Tissues are exposed to 1% Triton X-100 for 5 hrs. 每批组织的5小时存活率必须超过35%，由MTT测定(参见bbin体育官方网站表皮™ET-50方案)确定.
不育: 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.
病原体筛查: All cells are screened and are negative for HIV, hepatitis B and hepatitis C using PCR. 然而，没有一种已知的测试方法可以完全保证细胞是无病原体的. 因此, 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.
不合格批件通知: 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 our QC and sterility testing is done post-shipment, notification will be made as soon as possible (Under normal circumstances, % viability failures will be notified by Wednesday 5 p.m.; sterility failures will be notified within 8 days of shipment).
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