title: Establishment and optimization of NMR-based cell metabonomics study protocols for neonatal Sprague-Dawley rat cardiomyocytes.
abstract: A quenching, harvesting, and extraction protocol was optimized for cardiomyocytes NMR metabonomics analysis in this study. Trypsin treatment and direct scraping cells in acetonitrile were compared for sample harvesting. The results showed trypsin treatment cause normalized concentration increasing of phosphocholine and metabolites leakage, since the trypsin-induced membrane broken and long term harvesting procedures. Then the intracellular metabolite extraction efficiency of methanol and acetonitrile were compared. As a result, washing twice with phosphate buffer, direct scraping cells and extracting with acetonitrile were chosen to prepare cardiomyocytes extracts samples for metabonomics studies. This optimized protocol is rapid, effective, and exhibits greater metabolite retention.
authors.affiliation: Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang, 110016, PR China; National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.affiliation: National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China.
authors.forename: Bo
authors.initials: B
authors.lastname: Sun
authors.affiliation: National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China.
authors.forename: Rong
authors.initials: R
authors.lastname: Gao
authors.affiliation: National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Liu
authors.affiliation: National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China.
authors.forename: Fangting
authors.initials: F
authors.lastname: Dong
authors.affiliation: Evaluation and Research Center for Toxicology, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 20 Dongdajie Street, Fengtai District, Beijing, 100071, PR China.
authors.forename: Haiqin
authors.initials: H
authors.lastname: Fang
authors.affiliation: Evaluation and Research Center for Toxicology, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, 20 Dongdajie Street, Fengtai District, Beijing, 100071, PR China.
authors.forename: Shuangqing
authors.initials: S
authors.lastname: Peng
authors.affiliation: Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang, 110016, PR China.
authors.forename: Famei
authors.initials: F
authors.lastname: Li
authors.affiliation: National Center of Biomedical Analysis, 27 Taiping Road, Beijing, 100850, PR China. Electronic address: yanxzhon@163.com.
authors.forename: Xianzhong
authors.initials: X
authors.lastname: Yan
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D055442:Metabolome / Q000502:physiology*; D055432:Metabolomics / Q000379:methods*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
keywords: Cardiomyocytes; Cell metabonomics; Cellular metabolites extract; NMR