Microscale Gene Expression Analysis of Tumor-Associated Macrophages

Kuldeep S. Attri, Kamiya Mehla, Surendra K. Shukla, Pankaj Singh

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Macrophages, apart from being the key effector cells of the innate immune system, also play critical roles during the development and progression of various complex diseases, including cancer. Tumor-associated macrophages, infiltrate tumors during different stages of cancer progression to regulate motility, invasion, and intravasation to metastatic sites. Macrophages can exist in different polarization states associated with unique function in tumors. Since tumor-associated macrophages constitute a very small proportion of tumor cells, analysis of gene expression pattern using normal extraction buffer-based methods remains a challenging task. Therefore, it is imperative to develop low-throughput strategies to investigate transcriptional regulations from a small number of immune cells. Here, we describe an efficient, sensitive, and cost-effective approach for gene expression analysis of a small number of fluorescence-activated sorted tumor-associated macrophages. Our analyses from the different number of stable, primary, and sorted macrophages suggest 5,000 cells is an optimal number for performing quantitative, real-time PCR analysis of multiple genes. Our studies could detect expression of macrophage-specific genes from cultured primary macrophages, and FACS-sorted macrophages from different biological tissues without introducing biases in comparative gene expression ratios. In conclusion, our kit-based method for quantitative gene expression analysis from a small number of cells found in biological tissues will provide an opportunity to study cell-specific, transcriptional changes.

Original languageEnglish (US)
Article number2408
JournalScientific Reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

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Macrophages
Gene Expression
Neoplasms
Cell Count
Gene Expression Profiling
Genes
Real-Time Polymerase Chain Reaction
Immune System
Buffers
Fluorescence
Costs and Cost Analysis

ASJC Scopus subject areas

  • General

Cite this

Microscale Gene Expression Analysis of Tumor-Associated Macrophages. / Attri, Kuldeep S.; Mehla, Kamiya; Shukla, Surendra K.; Singh, Pankaj.

In: Scientific Reports, Vol. 8, No. 1, 2408, 01.12.2018.

Research output: Contribution to journalArticle

Attri, Kuldeep S. ; Mehla, Kamiya ; Shukla, Surendra K. ; Singh, Pankaj. / Microscale Gene Expression Analysis of Tumor-Associated Macrophages. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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