Transport Mode
Liquid Nitrogen Transportation
1、Adsorptive Liquid Nitrogen Transportation ,No free liquid nitrogen ,White vapor emission indicates normal operation.
2、Temperature monitoring devices should closely monitor the temperature inside the tank during transportation. If any abnormalities are detected, you can copy the PDF and Excel data from the temperature recorder. (One end of the temperature monitoring device can be unplugged to reveal a USB connector. Simply insert it into a computer to copy the data. If you encounter any difficulties, you can contact the sales staff and technical support for assistance.)
3、Alloy Combination Lock Designed to ensure no third-party access to the LN₂ tank or cell samples during transportation from dispatch to receipt, thereby safeguarding cell integrity.
4、GPS Tracker Enables real-time tracking of cell transportation routes to prevent loss.
5、Liquid Nitrogen Tank, temperature monitoring device, alloy combination lock, and GPS tracker are returnable components. Please store them properly and avoid damage; failure to comply will result in blacklisting.
Reference Citation
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Scope of Application
Mouse CD19-positive B cells are widely used in immunological research, including the establishment of models for autoimmune diseases, immunodeficiency, and B cell malignancies (such as leukemia and lymphoma); evaluating the efficacy of targeted therapies (e.g., CAR-T, antibody drugs); and investigating B cell development, signaling pathways, and regulatory mechanisms of immune function.
I 试剂与材料
Ø 冻存CD19阳性B细胞（Cat# LV-Bcell001/002）
Ø 培养基（完全RPMI1640，见下表）
Ø 37℃恒温水浴锅
Ø 生物安全柜
Ø 15ml离心管
Ø 37℃/5% CO2培养箱
Ø 宽口滴管和宽口移液枪头
Ø 移液枪
Ø 75%酒精
II 完全RPMI1640培养基
成分 | 用量 |
不完全RPMI1640溶液 | 500ml |
左旋谷氨酰胺（L-Glu） | 2mM |
2-巯基乙醇（2-ME） | 50μM |
青霉素(Penicillin) | 100U/ml |
链霉素(Streptomycin) | 100μg/ml |
FCS(56℃灭活30min) | 50ml |
特别提醒：复苏培养基中补加300mM葡萄糖，可提升细胞活力及活细胞数。
III 细胞的复苏与铺板
重要提示：解冻时，冻存细胞转移必须使用液氮转移；在整个复苏实验过程中必须全程使用宽口枪头。
1. 完全RPMI1640培养基放入37℃恒温水浴锅中让其充分预热。
2. 将冻存的细胞从冷藏位置迅速转移至37℃恒温水浴锅中。将冻存管尽可能多的浸入37℃水中，作规律的水平摇动，但必须确保冻存管盖子保持在水面以上。
3. 解冻冻存管至刚好完全溶解成液体，约90-120秒，。
4. 用75%酒精对冻存管消毒，并将其转移到生物安全柜。
5. 小心地将解冻后的细胞吸出，移至15ml离心管，并用培养基润洗一遍冻存管及吸取细胞的枪头。
6. 一边摇晃离心管，一边加入完全RPMI1640培养基至12ml（注意：前3ml要缓慢逐滴加入并摇晃，后面9ml可加快速度），最后上下颠倒混匀。
7. 20℃，650g，升速7，降速4，离心5min。
8. 弃上清，加入3ml完全RPMI1640培养基重悬细胞，并用台盼蓝染色计数（V细胞悬液：V0.4%台盼蓝=9：1，细胞活率使用血球计数板手工计数，勿用细胞计数仪；细胞总数使用细胞计数仪计数）测定细胞的活力、细胞总数，建议细胞总数检测3次，求取平均值；为节约时间，细胞活率检测1次。
9. 按实验要求的密度进行铺板。
在37℃/ 5% CO2培养箱中培养。
Thawed cells should not be refrozen, as repeated freezing may affect their function and status.
VI Regarding After-Sales Service
Quality Assurance & After-Sales Policy In the event of product quality issues, please: Collect original data Contact our sales representatives or technical support team immediately We will dispatch personnel to address the issue promptly.
After-Sales Validity Period & Submitted Original Data：
Resuscitation issues: Report any abnormalities immediately and provide results of Trypan blue staining or AO/PI staining.
Contamination issues: Report within 96 hours after resuscitation and provide phase-contrast microscope photos.
Purity issues: Report within one month of detection and provide immunofluorescence or flow cytometry results.
VII Contact Number
Company Telephone：0755-28284050
Technical Support：19902901483（ Dr. Zhou）
