Adipose Stem Cell Tissue Source Is Associated With Differential Expression Profiles In Cellular Mrna And Exosomal Mirna
Lauren Mangum1, Randolph Stone, II1, Shanmugasundaram Natesan1, Barbara Christy1, John Clifford2, Raina Kumar2, Andrew Cap1, Rasha Hammamieh2, Robert Christy1.
1US Army Institute of Surgical Research, Fort Sam Houston, TX, USA, 2US Army Center for Environmental Health Research, Fort Detrick, MD, USA.
Background: Severe traumatic injuries, including large total body surface area burns, limit the availability of autologous cells for wound repair. Adipose derived stem cells (ASCs) represent a potential source of autologous cells for use in reconstructive surgery and tissue engineering. We sought to better understand the role of ASC source on differential expression patterns of mRNA and miRNA to identify alterations to major signaling pathways. Methods: ASCs from subcutaneous adipose tissue associated with burned human (BH) or human abdominoplasty (HAP) skin were isolated under a protocol reviewed and approved by the Institutional Review Board. Commercially available ASCs were purchased from RoosterBio (RB). Cellular mRNA and exosomal miRNA were isolated from cell monolayers and culture media, respectively. Samples were submitted for sequencing and bioinformatics analysis. Significant differences in mRNA and miRNA expression was reported in log fold change at a threshold of p<0.05. Results: Comparing the cellular mRNA expression patterns, 202, 361, and 598 unique genes were identified for HAP, BH, and RB, respectively. When compared to RB, TGF-B, Beta-estradiol, and APP were identified as top upstream regulators for both HAP and BH. Analysis of exosomal contents revealed a small subset of differentially expressed miRNAs (6.5%) between HAP and BH groups. A larger subset of differentially expressed miRNAs was found between BH or HAP compared to RB ASCs (8.68% and 8.53%, respectively). Conclusions: Sequencing of mRNA and miRNAs from ASCs reveals source-dependent differences in several thousand genes governing diverse processes including cell cycle and proliferation. Analysis of mRNA and miRNA profiles indicated HAP and BH were more similar to each other than to RB. Preliminary analysis indicates tissue source, isolation procedures, and culturing conditions should be considered when developing stem cells or stem cell derived products for use in wound healing.
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