Department of Public Health Sciences, Henry Ford Health System, One Ford Place, Detroit, MI, 48202, USA.
Department of Biochemistry & Molecular Biology and RTSF/MSU Mass Spectrometry and Metabolomics Core, Michigan State University, 603 Wilson Road, Biochemistry Room 215, East Lansing, MI, USA.
Sci Rep. 2023 Mar 11;13(1):4063. doi: 10.1038/s41598-023-31287-3.
Methods for collection of placental tissue at room temperature for metabolic profiling are described. Specimens were excised from the maternal side of the placenta and immediately flash frozen or fixed and stored for 1, 6, 12, 24, or 48 h in 80% methanol. Untargeted metabolic profiling was performed on both the methanol-fixed tissue and the methanol extract. Data were analyzed using Gaussian generalized estimating equations, two sample t-tests with false discovery rate (FDR) corrections, and principal components analysis. Methanol-fixed tissue samples and methanol extracts had a similar number of metabolites (p = 0.45, p = 0.21 in positive vs. negative ion mode). In positive ion mode, when compared to flash frozen tissue, both the methanol extract and methanol-fixed tissue (6 h) had a higher number of metabolites detected (146 additional metabolites, p = 0.020; 149 additional metabolites, p = 0.017; respectively), but these associations were not found in negative ion mode (all p ≥ 0.05). Principle components analysis demonstrated separation of the metabolite features in the methanol extract, but similarity between methanol-fixed tissue and flash frozen tissue. These results show that placental tissue samples collected in 80% methanol at room temperature can yield similar metabolic data to flash frozen specimens.
描述了在室温下收集胎盘组织进行代谢组学分析的方法。从胎盘母体侧切取标本，立即进行快速冷冻或固定，并在 80%甲醇中储存 1、6、12、24 或 48 小时。对甲醇固定组织和甲醇提取物均进行非靶向代谢组学分析。使用高斯广义估计方程、具有错误发现率（FDR）校正的双样本 t 检验和主成分分析对数据进行分析。甲醇固定组织样本和甲醇提取物的代谢物数量相似（正离子模式下 p=0.45，负离子模式下 p=0.21）。与快速冷冻组织相比，在正离子模式下，甲醇提取物和甲醇固定组织（6 小时）检测到的代谢物数量更多（分别为 146 种额外的代谢物，p=0.020；149 种额外的代谢物，p=0.017），但在负离子模式下未发现这些关联（所有 p≥0.05）。主成分分析表明甲醇提取物中的代谢物特征分离，但甲醇固定组织和快速冷冻组织之间相似。这些结果表明，在室温下收集的 80%甲醇中的胎盘组织样本可以产生与快速冷冻标本相似的代谢数据。
