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 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

Peripheral Blood Mononuclear Cells (PBMC) It is an important tool for the study of the immune system. It has a wide range of scientific applications, including basic medicine, clinical research and drug development. Here is a summary of its core scientific uses:

1. Immunotherapy and oncology research

PBMC plays a central role in the development of immunotherapy drugs, mainly used to evaluate the killing ability of disease-free cells (such as T cells, NK cells) to tumor. For example:

Co-culture killing platform: The co-culture of PBMC and tumor cells, combined with flow cytometry assay (such as CSFE labeled target cells, CD45 labeled effector cells) to quantitatively analyze the specific killing effect of immune cells, for the development and optimization of CAR-T therapy and immune checkpoint inhibitors.

NK cell expansion and application: The isolation and expansion of highly active NK cells from PBMC are used in cancer immunotherapy research and drug screening, especially in in vitro models of solid and blood tumors.

2. Disease mechanisms and biomarkers research

PBMC provides key cellular models for unraveling disease mechanisms:

Heart and blood vessel disease: To study the pathologic mechanism and therapeutic target of patients with acute myocardial infarction by detecting the expression level of autophagy related proteins (such as LC3, Beclin1) in PBMC.

Metabolic diseases: To analyze lipid metabolism-related pathways in PBMC, such as STAT3, to reveal the molecular mechanisms of diseases such as nonalcoholic fatty liver disease (NAFLD).

Inflammation and Infection: To investigate the infection-free response of PBMC in viral (e.g., HIV, HBV) infection, or as a source of biomarkers for inflammatory diseases such as sepsis.

3. Drug Development and Metabolism Assessment

PBMC is used in drug development for efficacy evaluation, toxicity testing and metabolic studies:

In vitro metabolic model: Combined with liver microsomes and CYP450 enzymes, we evaluated phase I and phase II metabolic stability of drugs in PBMC and optimized drug design.

Drug Screening Platform: A PBMC- tumor co-culture system is used to screen candidate compounds that target epidemic-free regulation or anti-tumor activity, such as small molecule inhibitors or nanomedicines.

Toxic assessment: To detect the activation status of PBMC (e.g., inflammatory cytokine release, oxidative Stress response) and predict the side effects such as hepatic injury.

4. Regenerative Medicine and Personalized Medicine

PBMC has potential in cell therapy and disease model construction:

IPS cell reprogramming: Rapid construction of patient-specific iPS cell lines using cryopreserved PBMC for disease modeling (e.g., genetic diseases) and personalized drug testing.

Organoids and organochips: Integrate PBMC-derived disease-free cells into liver or tumor organoids to simulate immune microenvironment and study the interaction mechanism between disease-free cells and parenchymal cells.

5. Interdisciplinary and Translational Medicine Applications

The study of enterohepatic axis: To analyze the effect of intestinal microbial metabolites (such as LPS) on liver immunity through PBMC, to analyze the pathogenesis of enterogenous liver disease.

Single cell omics analysis: Combined with single cell sequencing technology, we analyzed the heterogeneity of gonopacytes and monocytes in PBMC, and revealed the disease-specific immune cell subsets.

summary

The use of PBMC covers immunotherapy, disease mechanism analysis, drug development, regenerative medicine and interdisciplinary research, and its versatility makes it an important bridge between basic research and clinical translation. In the future, with the development of single-cell technology and organoid models, the application of PBMC in precision medicine and personalized treatment will be further expanded.

Due to biannual updates of our product instruction manual, the following procedures are for reference only.

The most recent version included with the product shall prevail.

I 试剂与材料

Ø PBMC（Cat# LV-PBMC001/2/3）

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

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. 生物安全柜中，将10ml含300mM葡萄糖的完全RPMI1640培养基加入到15ml离心管中，37℃恒温水浴锅预热10min，然后转移到生物安全柜中；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PBMC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60-120秒。

4. 用75%酒精对冻存管消毒，并将其转移到生物安全柜。

5. 将解冻后的细胞轻轻吹打混匀2次，将细胞混匀后小心吸出，并以滴加方式转移至含已预热的复苏培养基的离心管中（注意：冻存管与枪头上残留细胞，可吸取1ml复苏培养基清洗冻存管与枪头并将其混入离心管的培养基中），轻微上下颠倒离心管2-3次混匀。

6. 离心力650g，升速7，降速4，离心5min。

7. 弃上清（倾倒去上清，保留底部残余液体，减少细胞损失），加入6-10ml完全RPMI1640培养基重悬细胞，并用台盼蓝染色和血球计数板计数（V_细胞悬液：V_{0.4%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。

8. 根据计数结果，按实验需求铺板。

9. 在37℃/ 5% CO₂培养箱中培养。

Post-Thaw Cells Must Not Undergo Refreezing.

IV 关于售后

如您发现产品有任何质量问题，请您收集原始数据，并第一时间联系公司销售或者技术支持，公司安排人员保证售后。每个实验室条件不同，操作人员习惯不同，熟练程度不一样，实验失败存在客观因素。如未严格按照说明书操作、超过售后时限，公司不做售后，请老师理解与支持。

售后有效期与提供的原始数据：

复苏问题：立刻报告异常；提供台盼蓝染色或者AO/PI染色。

污染问题：复苏96小时以内；提供相差显微镜照片。

纯度问题：一个月内；提供免疫荧光或者流式结果。

V 联系电话

公司电话：0755-28284050

技术支持：19902901483（周博士）