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小鼠乳腺癌类器官培养基
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
小鼠乳腺癌类器官培养基 (Mouse Breast Carcinoma Organoid Medium)， 是一种为促进小鼠乳腺癌类器官体外形成和扩增而开发的培养基。该产品是一种无菌的液体混合系统，含有维持目的细胞生长所必须的氨基酸、维生素、有机和无机化合物以及生长因子等。该培养基的配方可以提供一个合适的营养环境，选择性的促进体外小鼠乳腺癌类器官的生长和扩增。
使用说明
1. 获取肿瘤组织
(1) 按伦理规定处死小鼠后，完全切除肿瘤组织；
注：在这些步骤中采用最佳无菌条件可最大程度减少培养过程中的污染风险。为了便于操作，将小鼠仰卧固定在消毒的切割板或聚苯乙烯泡沫板上。用剪刀打开皮肤之前，给皮毛消毒。为了达到最佳的无菌效果，小心地将皮肤与腹壁分离，以防止腹膜穿孔。用新的剪刀，打开腹膜腔。
(2) 用镊子和剪刀取出肿瘤组织。将组织放入预冷的PBS中；
(3) 用冷的 PBS 轻轻清洗肿瘤组织2~3次；
(4) 加入适量预冷的PBS，尽量2~3小时内处理样本。
2、组织酶解消化
(1) 将装有组织的容器表面喷洒75%酒精消毒后放入生物安全柜，取组织于培养皿中，组织清洗液冲洗2次，转移至另一培养皿中用无菌手术刀切割至糊状，切割块大小约为1 mm³，加入5 mL组织清洗液，切割后的组织转移至15 mL离心管，1500 rpm 常温离心4 分钟。
(2) 弃上清，根据原组织块大小加入适当体积的 (消化液体积需是原组织体积的25~50倍)组织消化酶Ⅱ(T1521150)置于37 ℃振荡培养箱中进行组织消化，250~3000 rpm，一般消化20-60分钟。
注意:在此操作过程中须实时监测消化过程，因为过度消化可能会显著降低类器官形成效率。组织块出现絮状拉丝或组织块大小较原始体积减少一半时即可中止消化。
(3) 在消化完成的组织悬液中加入5~6 mL DF12 (含10%FBS)以终止消化，1500 rpm常温离心4分钟弃上清。
(4) 向细胞沉淀加入组织清洗液5 mL左右，重悬细胞沉淀，所获得悬液用100 μm筛网过滤，未消化完全组织可加入肿瘤组织消化酶Ⅱ(T1521150)再次消化，即重复步骤2-4，将两次得到的细胞沉淀合并。
(5) 观察是否有血细胞，若有血细胞，加入添加3~5 mL红细胞裂解液，冰上放置5 ~10分钟，1500 rpm、室温离心5分钟，弃上清。
(6) 加入1~5 mL类器官培养基重悬细胞，备用。
(7) 细胞计数：取重悬的细胞悬液12 µL与12 µL台盼蓝染液等体积充分混合后，取20 μL加入细胞计数板，选取三个不同视野计数观察细胞活率和细胞密度。
(8) 按照终浓度5~8×10⁵cells /mL细胞密度，取细胞悬液与基质胶等体积混匀，制备混合液，冰上操作。
注意：混匀动作要轻柔，避免产生气泡，若产生气泡，则混匀后冰上放置2~5分钟待气泡上浮后方可进行点胶。
(9) 取混合液50~60 μL点胶于24孔培养板(37 ℃提前预热)中，点胶完成后，培养板表面75%酒精消毒后，放置5% CO₂培养箱凝固30~60分钟直至胶体完全凝固。
注意：点胶尽量较少附着面积。
(10) 胶体完全凝固后，沿壁缓慢每孔加入700 μL类器官培养基，培养板表面75%酒精消毒后，放置37 ℃、5% CO₂培养箱培养。
(11) 镜下观察类器官形成情况；粒径大于50 μm即认为类器官已经形成，每2~3天更换一次培养基。
注意事项
1. 本产品使用前需平衡至室温。
2. 使用产品时应注意无菌操作，避免污染。
3. 本产品中含有抗生素，如无特别需要，不用额外添加其他抗生素，可直接使用。
4. 为保持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. 请于保质期内使用本产品，超出保质期后将无法达到理想的使用效果。
6. 本产品仅用于科研用途，不能用于体外诊断。
Mouse Breast Carcinoma Organoid Medium is a medium developed to promote the in vitro formation and expansion of mouse breast carcinoma organoids. This product is a sterile liquid mixture system containing essential amino acids, vitamins, organic and inorganic compounds, growth factors, and other components necessary for the growth of target cells. The formulation of this medium provides a suitable nutritional environment and selectively promotes the in vitro growth and expansion of mouse breast carcinoma organoids.
Instructions for Use
1. Tumor Tissue Collection
(1) Euthanize the mice in accordance with ethical regulations, then completely resect the tumor tissue.Note: Adopting optimal aseptic conditions during these steps minimizes the risk of contamination during culture. For ease of handling, fix the mouse in a supine position on a sterile dissection board or polystyrene foam board. Disinfect the fur before cutting open the skin with scissors. For optimal asepsis, carefully separate the skin from the abdominal wall to prevent peritoneal perforation. Open the peritoneal cavity using a new pair of scissors.
(2) Extract the tumor tissue using forceps and scissors. Place the tissue in pre-chilled PBS.
(3) Gently wash the tumor tissue 2-3 times with cold PBS.
(4) Add an appropriate volume of pre-chilled PBS and process the sample within 2-3 hours if possible.
2. Enzymatic Tissue Digestion
(1) Disinfect the surface of the container holding the tissue by spraying 75% alcohol, then place it in a biological safety cabinet. Transfer the tissue to a culture dish, rinse twice with tissue washing solution, then transfer to another culture dish and mince into a paste-like state with a sterile scalpel, to a fragment size of approximately 1 mm³. Add 5 mL of tissue washing solution, transfer the minced tissue to a 15 mL centrifuge tube, and centrifuge at 1500 rpm at room temperature for 4 minutes.
(2) Discard the supernatant. Add an appropriate volume of Tissue Digestion Enzyme II (T1521150) according to the size of the original tissue block (digestion solution volume should be 25-50 times the volume of the original tissue). Digest the tissue in a 37 °C shaking incubator at 250-3000 rpm for a general digestion time of 20-60 minutes.
Note: The digestion process must be monitored in real time, as over-digestion may significantly reduce organoid formation efficiency. Digestion may be terminated when the tissue mass becomes flocculent with filamentous stretching or is reduced to half its original volume.
(3) Add 5-6 mL of DF12 (containing 10% FBS) to the digested tissue suspension to terminate digestion. Centrifuge at 1500 rpm at room temperature for 4 minutes and discard the supernatant.
(4) Add approximately 5 mL of tissue washing solution to the cell pellet to resuspend the cells. Filter the resulting suspension through a 100 μm mesh filter. Undigested tissue may be re-digested with Tumor Tissue Digestion Enzyme II (T1521150) by repeating Steps 2-4, then combine the cell pellets obtained from both digestions.
(5) Check for the presence of blood cells. If present, add 3-5 mL of red blood cell lysis buffer, place on ice for 5-10 minutes, centrifuge at 1500 rpm at room temperature for 5 minutes, and discard the supernatant.
(6) Add 1-5 mL of organoid medium to resuspend the cells; set aside for later use.
(7) Cell counting: Mix 12 µL of resuspended cell suspension with an equal volume of 12 µL trypan blue staining solution thoroughly. Pipette 20 μL into a cell counting chamber, and count cell viability and density in three random fields of view.
(8) At a final cell density of 5-8×10⁵ cells/mL, mix the cell suspension with an equal volume of matrix gel on ice to prepare the mixture.
Note: Mix gently to avoid air bubbles. If bubbles form, place the mixture on ice for 2-5 minutes to allow bubbles to rise before seeding.
(9) Pipette 50-60 μL of the mixture per dome onto a 24-well culture plate (pre-warmed at 37 °C). After seeding, disinfect the plate surface with 75% alcohol, then place in a 5% CO₂ incubator for 30-60 minutes until the gel is fully solidified.
Note: Minimize the attachment area of the gel domes during seeding.
(10) After complete gel solidification, slowly add 700 μL of organoid medium per well along the inner wall. Disinfect the plate surface with 75% alcohol, then culture at 37 °C in a 5% CO₂ incubator.
(11) Observe organoid formation under a microscope. Organoids are considered formed when their diameter exceeds 50 μm. Replace the medium every 2-3 days.
Precautions
1. This product should be equilibrated to room temperature before use.
2. Practice aseptic technique during use to avoid contamination.
3. This product contains antibiotics. Unless specifically required, no additional antibiotics need to be supplemented; it can be used directly.
4. To maintain optimal performance, do not leave this product at room temperature or elevated temperatures overnight.
5. Use this product within its expiration date. Expired product may not achieve the desired performance.
6. This product is for research use only and is not intended for in vitro diagnostic procedures.
规格
别名
小鼠乳腺癌类器官生长培养基
英文别名
Mouse Breast Carcinoma Organoid Growth Medium | Mouse Breast 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
Mouse Breast 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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通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0535265 |
分析证书 | M1521146 |
| ZJ26F0535264 |
分析证书 | M1521146 |
技术文档和文章
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