Dataset Information

Derivatized heparin hexaccharides LC-MS/MS

ABSTRACT: We have developed a novel chemical derivatization method coupled tandem mass spectrometry to resolve the structural sequences of sulfated glycosaminoglycans. In order to evaluate and demonstrate our method capable for native sulfated oligosaccharides, a complex heparin hexasaccharide was used.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

SUBMITTER: Hao Liu

LAB HEAD: Joshua S. Sharp

PROVIDER: PXD021263 | Pride | 2020-09-21

REPOSITORIES: Pride

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Publications

Peracylation Coupled with Tandem Mass Spectrometry for Structural Sequencing of Sulfated Glycosaminoglycan Mixtures without Depolymerization.

Liu Hao H Liang Quntao Q Sharp Joshua S JS

Journal of the American Society for Mass Spectrometry 20200918 10

The structures of glycosaminoglycans (GAGs), especially the patterns of modification, are crucial to modulate interactions with various protein targets. It is very challenging to determine the fine structures using liquid chromatography-mass spectrometry (LC-MS) due in large part to the gas-phase sulfate losses upon collisional activation. Previously, our group reported a method for fine structure analysis that required permethylation of the GAG oligosaccharide. However, uncontrolled depolymeriz ...[more]

PMID: 32902282

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Project description:Background: Mesenchymal stromal cells (MSCs) are a promising cell source for tissue engineering and regenerative medicine. In our lab, we found that cell preparations from bone marrow of many donors had a limited capacity for in vitro-differentiation into the osteogenic and chondrogenic lineages although this is a capacity claimed to be inherent to this kind of cells. Therefore the current study was designed to test the hypothesis whether the amount of heparin used as anti-coagulant during bone marrow harvest has inhibitory influence on the in vitro-differentiation capacity of the isolated MSCs. Methods: Bone marrow was obtained from twelve donors with good physical condition during preparation of the femoral cavity for the implantation of the femoral stem for a total hip arthroplasty in the absence or presence of heparin. Isolated MSCs were characterized by plastic adhesion, morphology, population doubling times, expression of cell surface antigens and in vitro-differentiation into the adipogenic, osteogenic and chondrogenic lineages. In addition, transcriptome analyses were performed. Results: Bone marrow prepared in the absence of heparin showed a comparable processability as bone marrow in the presence of heparin. Notably, no coagulation was observed. The number of mononuclear cells retrieved from the density gradient was independent of heparin addition. Moreover, none of the phenotypic or functional parameters assessed correlated with the addition of heparin. Transcriptome and quantitative realtime PCR analyses demonstrated that some selected genes were up- or downregulated in the cells isolated with the addition of heparin. However, this statement held true for only some but not all of the donors investigated in the present study. Conclusions: No correlation with heparin addition was observed with respect to the parameters assessed in the present study. This implies the absence of consequences both for the results of in vitro-investigations and also during in vivo-application, thereby ruling out heparin as a potential source of disparate results that are described in the literature.

Dataset Information

Derivatized heparin hexaccharides LC-MS/MS

Publications

Peracylation Coupled with Tandem Mass Spectrometry for Structural Sequencing of Sulfated Glycosaminoglycan Mixtures without Depolymerization.

Similar Datasets

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