Evaluating the Effect of Cell Culture on Gene Expression in Primary Tissue Samples Using Microfluidic-Based Single Cell Transcriptional Analysis

Microarrays

Michael Januszyk²  Robert C. Rennert²  Michael Sorkin²  Zeshaan N. Maan²  Lisa K. Wong²  Alexander J. Whittam²  Arnetha Whitmore²  Dominik Duscher²  Geoffrey C. Gurtner¹ 

[1] Department of Surgery, Division of Plastic and Reconstructive Surgery, 257 Campus Drive West, Hagey Building GK-201, Stanford, CA 94305-5148, USA;

关键词: microfluidics;    singe-cell;    transcriptional analysis;    gene expression;    bioinformatics;    subpopulations;    cell culture;   

DOI  :  10.3390/microarrays4040540

来源: mdpi

【 摘 要 】

Significant transcriptional heterogeneity is an inherent property of complex tissues such as tumors and healing wounds. Traditional methods of high-throughput analysis rely on pooling gene expression data from hundreds of thousands of cells and reporting a population-wide average that is unable to capture differences within distinct cell subsets. Recent advances in microfluidic technology have permitted the development of large-scale single cell analytic methods that overcome this limitation. The increased granularity afforded by such approaches allows us to answer the critical question of whether expansion in cell culture significantly alters the transcriptional characteristics of cells isolated from primary tissue. Here we examine an established population of human adipose-derived stem cells (ASCs) using a novel, microfluidic-based method for high-throughput transcriptional interrogation, coupled with advanced bioinformatic analysis, to evaluate the dynamics of single cell gene expression among primary, passage 0, and passage 1 stem cells. We find significant differences in the transcriptional profiles of cells from each group, as well as a considerable shift in subpopulation dynamics as those subgroups better able to adhere and proliferate under these culture conditions gradually emerge as dominant. Taken together, these findings reinforce the importance of using primary or very early passage cells in future studies.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.