Human Non-Small Cell Lung Carcinoma (NSCLC) Peripheral Blood Mononuclear Cells (PBMCs)

Description

About Non-small Cell Lung Carcinoma (NSCLC) and Our Donors

Non-small cell lung carcinoma (NSCLC) makes up approximately 85% of all lung cancers and is characterized by transformed lung epithelium. The most common types of NSCLC are adenocarcinoma, large cell carcinoma, and squamous cell carcinoma. Lung adenocarcinoma is the most common lung cancer among non-smokers, while squamous cell carcinoma is the most closely associated type of lung cancer with smokers. Large cell carcinoma, meanwhile, encompasses a heterogeneous group of malignant neoplasms that are undifferentiated from lung epithelium.

The leading cause of lung cancer is smoking, in which thousands of compounds in cigarette smoke promote DNA damage. In a healthy organism, cells have mechanisms that repair damaged DNA. However, if not repaired, the mutated DNA can give rise to cancerous cells. In NSCLC, studies have shown that at least nine DNA repair genes are silenced, thereby preventing the DNA damage repair machinery from being expressed and operational. Coupled with the DNA damage incurred by smoke, a high rate of mutated DNA goes unrepaired, resulting in cancerous cells.

Besides silencing of DNA repair genes, mutations in certain genes have also been found to promote NSCLC. Approximately 10-35% of NSCLC patients have mutations in the EGFR gene, which can be treated with tyrosine kinase inhibitors, such as gefitinib and erlotinib, to some success. Similarly, almost 7% of NSCLC patients have EML4-ALK translocations or mutations in the ROS1 gene. Here, treatment with ALK inhibitors may provide some benefit to patients.

While the tyrosine kinase and ALK inhibitors have been important treatments for some NSCLC patients, recent approvals for checkpoint protein inhibitors provide an additional treatment option for patients that do not have mutations in the EGFR or ALK gene. Together, these observations give researchers better insight into NSCLC and development of therapies to treat it.

Our PBMCs from NSCLC patients are sourced from a renowned, world-class blood collection center that has been a pioneer in many screening processes. All PBMCs have been tested for various infectious diseases, including HIV and Hepatitis B/C, as well as normal levels of hemoglobin. Further, donors undergo a screening questionnaire to disclose vital statistics, medication, cancer history, and travel history to ensure that their PBMCs will reliably serve your research endeavors.

Application Summary for PBMCs from NSCLC Patients

PBMCs from NSCLC patients can be used for a wide variety of immunology-based applications, such as ex-vivo applications that characterize immune cell populations based on immunophenotyping. These PBMCs are also an ideal source of effector cells to perform functional experiments with, such as those that interrogate cytotoxicity or cytokine production, to determine if lymphocytes from NSCLC patients have alterations in immune function.

Isolation of specific immune cell types from PBMCs of NSCLC patients can also be performed to obtain an enriched population in order to carry out genome or transcriptome sequencing to determine if NSCLC-driven alterations are present.

Isolation of PBMCs from NSCLC Patients

Peripheral blood is obtained from responsible facilities that have many years of experience collecting samples. Since the facilities are local, the blood is quickly transported to iQ Biosciences’ laboratory for processing.

Once at our lab, PBMCs from NSCLC patients are isolated by performing density centrifugation with Ficoll, a high molecular weight sucrose solution, to remove red blood cells under sterile conditions. Depending on the species (human or non-human) of blood, either 1.077 or 1.084 g/ml Ficoll is used, resulting in a perfect layer of mononuclear cells that are separated from plasma, platelets, granulocytes, and erythrocytes. Additional quality control steps are taken to isolate and prepare PBMCs that ensure the highest viability for cryopreservation and downstream experimental applications.

Cryopreservation and Storage

Our PBMCs from NSCLC patients are cryopreserved carefully using iQ Biosciences’ cryopreservation protocol that ensures high viability (> 90%) after thawing.

Cells should be stored at < -120°C once they are received, such as within a liquid nitrogen tank (vapor phase).

Quality Control Process

We implement random sampling per lot to test for viability and cell counts to ensure they meet specifications, which are recorded on the Certificate of Analysis that is included with each shipment. Cell counts are obtained using a manual hemocytometer and then cross-referenced with an automated cell counter. Each lot is also characterized for unique cell populations by immunophenotyping, in which the results are also recorded on the Certificate of Analysis.