本产品用于组织前处理过程中清洗肿瘤或者正常组织,其中针对性的添加多种抗生素进行组合,发挥协同抗菌作用,可以有效降低原代细胞培养过程中的污染率。
使用说明
1. 提前将消化酶从-20℃冰箱取出置于2-8℃融化;
2. 使用消化酶稀释液 (货号D1521149)按照1: 1比例稀释组织消化酶 (货号O1521148),配制成工作浓度;
3. 将组织转移至培养皿中,加入3 mL组织清洗液冲洗组织,反复2-3次;
4. 在无菌条件下将组织机械切割成1~2 mm³大小之后转移至15 mL无菌离心管中;
5. 加入5 mL组织清洗液,1500 rpm常温离心3 min;
6. 弃上清,加入12 mL工作浓度的消化酶,标记样本名称及编号并封口,放入恒温振荡培养箱中消化,设置参数37 ℃、200~300 rpm (根据样本质地设置转速),待组织消化呈团絮状,没有明显肉眼可见硬块终止消化;
7. 取出消化好的样本,1500 rpm室温离心3 min。
8. 弃上清后,加入6 mL含血清培养基进行终止消化,1500 rpm室温离心3 min。
9. 弃上清后加入原代细胞培养基重悬后待用。
注意事项
1. 使用产品时应注意无菌操作,避免污染。
2. 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。
3. 本产品仅用于科研用途,不能用于体外诊断。
This product is used for washing tumor or normal tissues during tissue pretreatment. It contains a combination of multiple antibiotics to exert synergistic antibacterial effects, which can effectively reduce contamination rates in primary cell culture.
Instructions for Use
1. Take the digestive enzyme out of the -20℃ refrigerator in advance and thaw it at 2–8℃.
2. Dilute Tissue Digestion Enzyme (Catalog No. O1521148) at a ratio of 1:1 with Digestive Enzyme Diluent (Catalog No. D1521149) to prepare working solution.
3. Transfer tissues into a petri dish, add 3 mL tissue washing solution to rinse tissues, and repeat rinsing 2–3 times.
4. Under aseptic conditions, mechanically cut tissues into 1–2 mm³ fragments, then transfer them into sterile 15 mL centrifuge tubes.
5. Add 5 mL tissue washing solution and centrifuge at 1500 rpm at room temperature for 3 minutes.
6. Discard the supernatant, add 12 mL working-concentration digestive enzyme. Label sample names and numbers, seal the tubes, and place them in a constant-temperature shaking incubator for digestion. Set parameters at 37℃, 200–300 rpm (adjust speed according to tissue texture). Terminate digestion when tissues become flocculent with no visible hard lumps.
7. Take out digested samples and centrifuge at 1500 rpm at room temperature for 3 minutes.
8. Discard the supernatant, add 6 mL serum-containing medium to terminate digestion, and centrifuge at 1500 rpm at room temperature for 3 minutes.
9. Discard the supernatant, resuspend cells with primary cell medium for subsequent use.
Precautions
1. Strict aseptic operation is required during use to avoid contamination.
2. Use the product within the shelf life. Ideal experimental results cannot be guaranteed after expiration.
3. For research use only, not for in vitro diagnosis.