计算溶液所需的质量、体积或浓度。
人鼻咽癌类器官培养基
有货
为什么选择此级别
BioReagent,无菌,用于细胞培养 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
人鼻咽癌类器官培养基 (Human Nasopharyngeal Carcinoma Organoid Medium)是一种为促进人源鼻咽癌类器官体外形成和扩增而开发的培养基。它是一种无菌的液体混合系统，含有必需和非必需的氨基酸、维生素、有机和无机化合物、生长因子、微量矿物质等。该培养基的独特配方可以提供一个最佳人源鼻咽癌类器官体外培养环境，选择性地促进体外人源鼻咽癌类器官的生长。
使用说明
1. 实验前准备：
(1) 15 mL无菌离心管、1.5 mL离心管、24孔细胞培养板、移液器、移液管、无菌枪头等表面消毒后放入生物安全柜中紫外照射30 min。
(2) 提前30 min从4 ℃冰箱取出鼻咽癌类器官培养基平衡至室温，提前1 h从-20 ℃冰箱取出所需体积的基质胶冰上融化。
2. 鼻咽癌类器官培养
(1) 将获取的鼻咽癌原代细胞悬液计数，根据计算好的体积吸取细胞悬液加入预冷的1.5 mL离心管中。
(2) 补齐预冷的培养基至所需体积，将预冷后的细胞悬液加入等体积的基质胶中轻轻混匀 (全程冰上操作)，使得鼻咽癌原代细胞终浓度为5×10⁶个细胞/毫升。
(3) 然后将细胞与基质胶的混合液按照50 μL/孔呈半球形点胶于24孔板。
(4) 将培养板放入5% CO₂、37 ℃培养箱中30 min，待基质胶完全凝固。
(5) 沿孔壁每孔缓慢加入500 μL预先恢复至室温的类器官培养基，置5% CO₂、37 ℃培养箱培养。
(6) 3天后镜下观察类器官形成情况，粒径大于30~50 μm即认为类器官已经形成，每3天更换一次培养基进行培养，持续观察类器官大小，镜下观察大部分类器官大小均大于30~50 μm且大小不再增大即可收集类器官进行后续实验。
3. 鼻咽癌类器官收集
弃培养基，每孔加入500 μL预冷DF12，反复吹打至基质胶完全脱离板底，收集至15 mL离心管中，1500 rpm离心5 min，轻轻吸去上清和基质胶层，待用。
4. 鼻咽癌类器官传代
(1) 收集的类器官，加入3-5 ml类器官解离试剂 (O1521153)，37 ℃，200 rpm水平摇床振荡消化(具体时间根据类器官大小和数量决定，以肉眼可见颗粒明显减小一半为准，可中间取消化的类器官显微镜观察，类器官消化至包含3~10个细胞团块最佳)。
(2) 消化完全后1500 rpm离心3 min，弃上清，使用清洗液重悬后1500 rpm离心3 min，加入预冷的类器官培养基进行重悬，计数，按照培养步骤操作继续培养。
注意事项
1. 本产品在使用前需平衡至室温。
2. 样本需为肿瘤组织，且肿瘤细胞比例越高 (>50%),类器官培养成功率越高。
3. 使用产品时应注意无菌操作，避免污染。
4. 为保持本产品的最佳使用效果，不宜将其过夜放置于室温或较高的温度环境中。
5.类器官培养过程要轻柔，避免基质胶凝固后散胶。
6. 请于保质期内使用本产品，超出保质期，必须放弃使用。
7. 本产品仅用于科研用途，不能用于体外诊断。
Human Nasopharyngeal Carcinoma Organoid Medium is a medium developed to promote the in vitro formation and expansion of human nasopharyngeal carcinoma organoids. It is a sterile liquid mixture system containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, growth factors, trace minerals, etc. The unique formulation of this medium provides an optimized in vitro culture environment for human nasopharyngeal carcinoma organoids and selectively promotes their growth in vitro.
Instructions for Use
1. Pre‑experiment Preparation
(1) Disinfect the surfaces of 15 mL sterile centrifuge tubes, 1.5 mL centrifuge tubes, 24‑well cell culture plates, pipettes, pipettors, sterile pipette tips, etc., then place them in a biological safety cabinet and expose to UV irradiation for 30 min.
(2) Remove the nasopharyngeal carcinoma organoid medium from the 4 °C refrigerator 30 min in advance and equilibrate to room temperature. Take the required volume of matrix gel from the −20 °C refrigerator and thaw on ice for 1 h in advance.
2. Culture of Nasopharyngeal Carcinoma Organoids
(1) Count the obtained primary nasopharyngeal carcinoma cell suspension, and transfer the calculated volume of cell suspension into a pre‑cooled 1.5 mL centrifuge tube.
(2) Top up with pre‑cooled medium to the required volume. Add the pre‑cooled cell suspension to an equal volume of matrix gel and mix gently (all operations on ice) to achieve a final concentration of primary nasopharyngeal carcinoma cells of 5 × 10⁶ cells/mL.
(3) Then dispense 50 μL of the cell‑matrix gel mixture per well into a 24‑well plate in a hemispherical dome.
(4) Place the culture plate in a 37 °C incubator with 5% CO₂ for 30 min until the matrix gel is fully solidified.(5) Slowly add 500 μL of room‑temperature equilibrated organoid medium along the inner wall of each well, and culture in a 37 °C incubator with 5% CO₂.
(6) Observe organoid formation under a microscope after 3 days. Organoids are considered formed when their diameter exceeds 30-50 μm. Replace the medium every 3 days and continue monitoring organoid size. When most organoids are larger than 30–50 μm and no longer increase in size under microscopy, collect organoids for subsequent experiments.
3. Harvesting of Nasopharyngeal Carcinoma Organoids
Discard the medium. Add 500 μL of pre‑cooled DF12 to each well, and pipette repeatedly until the matrix gel is completely detached from the plate bottom. Collect the suspension into a 15 mL centrifuge tube, centrifuge at 1500 rpm for 5 min, gently aspirate the supernatant and matrix gel layer, and set aside for later use.
4. Subculture of Nasopharyngeal Carcinoma Organoids
(1) Add 3-5 mL of Organoid Dissociation Reagent (O1521153) to the collected organoids. Digest with horizontal shaking at 37 °C, 200 rpm (actual digestion time depends on organoid size and quantity, until visible particles are obviously reduced by half. Midway microscopic observation is recommended; optimal digestion is achieved when organoids are dissociated into clusters of 3-10 cells).
(2) After complete digestion, centrifuge at 1500 rpm for 3 min and discard the supernatant. Resuspend with washing solution, centrifuge again at 1500 rpm for 3 min, then resuspend the pellet in pre‑cooled organoid medium. Count and continue culture following the seeding procedure.
Precautions
1. This product should be equilibrated to room temperature before use.
2. Samples should be tumor tissue; a higher tumor cell proportion (>50%) results in a higher success rate of organoid culture.
3. Practice aseptic technique during use to avoid contamination.
4. To maintain optimal performance, do not leave this product at room temperature or elevated temperatures overnight.
5. Handle gently during organoid culture to prevent disruption of the solidified matrix gel.
6. Use this product within its expiration date. Discard any expired product.
7. This product is for research use only and is not intended for in vitro diagnostic procedures.
规格
别名
人鼻咽癌类器官生长培养基
英文别名
Human Nasopharyngeal Carcinoma Organoid Growth Medium | Human Nasopharyngeal Carcinoma Organoid Culture Media
规格或纯度
BioReagent, 用于细胞培养, 无菌
稳定性与储存
Store at -20℃ long term (12 months).
英文名称
Human Nasopharyngeal Carcinoma Organoid Medium
储存条件
-20°C储存
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0535269 |
分析证书 | N1521145 |
| ZJ26F0535268 |
分析证书 | N1521145 |
技术文档和文章
溶液计算器
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