Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Deja Clay - University of Nevada, Las Vegas
Co-Author(s): Yingjia Chen and Dr. Tai Guo, University of Georgia, Athens, Georgia
Tumor-associated macrophages (TAMS) are one of many immune cells shown to induce tumorigenesis and metastasis. Mouse models have been and continue to be used to examine the cross-talk between the tumor microenvironment and cells that promote tumor growth. Nevertheless, several protocols have been designed to guide researchers on how to accurately isolate macrophages and other immune cells from tumors. While each method yields an adequate quantity of cells, an assessment on which protocol produces the most has yet to be investigated. We conducted experiments using three different protocols on three groups to determine which yield the highest number or percentage of immune cells. Prostate tumors were harvested from C57BL6 mice and the percentages of CD8+, CD4+, and macrophages were compared between each protocol. Protocol 1 only involved the laceration of the prostate tumor. Protocol 2 and 3 underwent the same step; however, two additional procedures (enzyme digestion & red blood cell lysis) were completed on these protocols. Protocol 3 was the only method to include one-step double Ficoll gradient centrifugation. Overall, Protocol 1 isolated the largest percentage of CD8+, CD4+, and macrophages. It was hypothesized that enzymatic digestion performed during Protocols 2 and 3 may have resulted in the loss of cell surface receptors, which could explain the lower percentage in Groups 2 and 3.Not Submitted
Funder Acknowledgement(s): NSF HBCU-UP Targeted Infusion Project, Award No. 1533498
Faculty Advisor: Dr. Tai Guo, email@example.com
Role: I aided in collection of the prostate tumors, lacerating the tumors, and analyzing the data collected.