Laser Capture Microdissection of Epithelium from a Wound Healing Model for MicroRNA Analysis
MicroRNAs are ~22 nucleotide-long noncoding RNAs influencing many cellular processes (including wound healing) by their regulatory functions on gene expression. The ability to analyze microRNA in different cells at the wound site is essential for understanding the critical role(s) of microRNA during various phases of wound healing. Laser capture micro-dissection (LCM) is an effective method to distinguish between relevant and non-relevant cells or tissues and enables the researcher to obtain homogeneous, ultra-pure samples from heterogeneous starting material. We present here our protocol for procuring epithelial cells from a mouse wound healing model using a Leica LMD7000 Laser Microdissection system, as well as the RNA isolation and downstream microRNA analysis. Using this method, researchers can selectively and routinely analyze regions of interest down to single cells to obtain results that are relevant, reproducible, and specific.
Key wordsLCM MicroRNA Wound healing RNA isolation Epithelial cells Mouse Frozen sample
This work was supported in part by NIH PHS grants (R21DE025926, R03CA171436, R01GM50875, and S10RR026493) and a Lilly USA Research Award in Cancer Prevention and Early Detection. Dr. Alyne Simões was supported by a scholarship from São Paulo Research Foundation (2016/16332-0). We thank Dr. Wendy Cerny for editorial assistance.
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