Cryopreservation and Gene Expression Analysis of PBMCs

Patients’ peripheral blood was centrifuged in Vacutainer CPT tubes (BD) at 1,800×g/18°C for 20 min with slow acceleration/deceleration. PBMCs were suspended in CryoStor (BioLife Solutions/CS10) freezing media and stored in liquid nitrogen [47 (link)]. For gene expression analysis, mRNA was isolated from PBMCs from baseline and the second week draw during CMT from 4 HPV⁺ patients and 2 HPV^– patients (number 7, 8, 11, 12, 13, and 14) (Table 1) using mRNA isolation kit on Maxwell Rapid Sample Concentrator instrument (Promega). RNA quality was verified using the Bioanalyzer-2100 (Agilent). Gene expression was analyzed using Human PanCancer Immune Profiling panel (XT-CSO-HIP1-12) on the nCounter system (NanoString Technologies, WA) following manufacturer’s recommendations. Data analysis was done using nSolver 3.0 software (NanoString Technologies, WA). Raw data were normalized by stable housekeeping gene selected automatically by the system (Supplementary Table 6). Heat-maps and cell type profiling analysis were generated by the nSolver advanced analysis.

Free full text: Click here

Sampath S., Won H., Massarelli E., Li M., Frankel P., Vora N., Vora L., Maghami E, & Kortylewski M. (2018). Combined modality radiation therapy promotes tolerogenic myeloid cell populations and STAT3-related gene expression in head and neck cancer patients. Oncotarget, 9(13), 11279-11290.

Publication 2018

Vacutainer CPT tubes CryoStor Maxwell Rapid Sample Concentrator instrument Bioanalyzer-2100 nCounter system nSolver 3.0 software

Acceleration Blood Cell type Deceleration G 800 Gene expression Gene expression analysis Hip1 Housekeeping gene Human Isolation Maps Mrna Nitrogen Patients Promega

Corresponding Organization : City Of Hope National Medical Center

Top 5 similar protocols

Variable analysis

independent variables

Patients' peripheral blood centrifugation conditions (1,800×g/18°C for 20 min with slow acceleration/deceleration)

dependent variables

Gene expression in PBMCs from baseline and the second week draw during CMT

control variables

PBMC samples from 4 HPV+ patients and 2 HPV- patients

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!