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• Original Article •
Extraction methods optimization in high-field MRS study of the human umbilical cord mesenchymal stem cells
DAI Hai-yang  HONG Bi-kai  XIAO Ye-yu  YANG Yu-ping  MA Lian  CHEN Yao-wen  WU Ren-hua 

DOI:10.3969/j.issn.1674-8034.2011.06.008.


[Abstract] Objective: To explore the suitable cell extraction methods that most fit in vitro MRS study by analyzing the extraction efficiencies and detecting sensitivity, and to make some optimizations to the choice of different extraction procedures.Materials and Methods: Human umbilical cord mesenchymal stem cells were cultured and collected. The water-soluble metabolites were extracted by perchloric acid (PCA) and the lipid-soluble metabolites were extracted by methanol-chloroform (M/C ), and we employed a dual phase extraction (DPE) procedure by methanol-chloroform-water to get both fractions from the same cell batch. The net weights from different extraction methods were compared and analyzed using Student paired t-test.Results: Compared with the net yields from DPE, the PCA extraction procedure has a higher efficiency in extracting water-soluble metabolites while M/C is efficient in extracting lipid-soluble metabolites. MRS is highly sensitive in detecting extremely small quantity of lipid-soluble metabolites.Conclusion: We can employ PCA combined with M/C to obtain both water- and lipid-soluble metabolites for the efficient use of cells in MRS analysis.
[Keywords] Mesenchymal stem cells；Magnetic resonance spectroscopy；Metabolite extraction；Optimization

DAI Hai-yang Department of Medical Imaging, The Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China

HONG Bi-kai Department of Medical Imaging, The Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China

XIAO Ye-yu Department of Medical Imaging, The Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China

YANG Yu-ping Department of pediatrics, The Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China

MA Lian Department of pediatrics, The Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China

CHEN Yao-wen Shantou University Central Laboratory, Shantou 515041, China

WU Ren-hua* Department of Medical Imaging, The Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China

*Correspondence to: Wu RH, rhwu@stu.edu.cn

Conflicts of interest   None.

Received  2011-07-12
Accepted  2011-09-25
DOI: 10.3969/j.issn.1674-8034.2011.06.008
DOI:10.3969/j.issn.1674-8034.2011.06.008.

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