Characterization of secretomes provides evidence for adipose-derived mesenchymal stromal cells subtypesReport as inadecuate




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Stem Cell Research and Therapy

, 6:221

First Online: 11 November 2015Received: 13 February 2015Revised: 18 February 2015Accepted: 20 October 2015

Abstract

IntroductionThis study was aimed at deciphering the secretome of adipose-derived mesenchymal stromal cells ADSCs cultured in standard and hypoxic conditions to reveal proteins, which may be responsible for regenerative action of these cells.

MethodsHuman ADSCs were isolated from 10 healthy donors and cultured for 3–4 passages. Cells were serum deprived and cell purity was assessed using multiple cell surface markers. Conditioned media was collected and analyzed using LC-MS with a focus on characterizing secreted proteins.

ResultsPurity of the ADSC assessed as CD90+-CD73+-CD105+-CD45-CD31- cells was greater than 99 % and viability was greater than 97 %. More than 600 secreted proteins were detected in conditioned media of ADSCs. Of these 100 proteins were common to all cultures and included key molecules involved in tissue regeneration such as collagens and collagen maturation enzymes, matrix metalloproteases, matricellular proteins, macrophage-colony stimulating factor and pigment epithelium derived factor. Common set of proteins also included molecules, which contribute to regenerative processes but were not previously associated with ADSCs. These included olfactomedin-like 3, follistatin-like 1 and prosaposin. In addition, ADSCs from the different subjects secreted proteins, which were variable between different cultures. These included proteins with neurotrophic activities, which were not previously associated with ADSCs, such as mesencephalic astrocyte-derived neurotrophic factor, meteorin and neuron derived neurotrophic factor. Hypoxia resulted in secretion of 6 proteins, the most prominent included EGF-like repeats and discoidin I-like domains 3, adrenomedullin and ribonuclease 4 of RNase A family. It also caused the disappearance of 8 proteins, including regulator of osteogenic differentiation cartilage-associated protein.

ConclusionsHuman ADSCs with CD90+-CD73+-CD105+-CD45-CD31-PDGFRβ+-NG2+-CD146+− immunophenotype secrete a large array of proteins, the most represented group is comprised of extracellular matrix components. Number of secreted proteins is largely unaffected by prolonged hypoxia. Variability in the secretion of several proteins from cultured ADSCs of individual subjects suggests that these cells exist as a heterogeneous population containing functionally distinct subtypes, which differ in numbers between donors.

KeywordsAdipose-derived mesenchymal stromal cells Secretome profiling Hypoxic response Regeneration Cell therapy AbbreviationsADSCsAdipose-derived mesenchymal stromal cells

BDNFBrain derived neurotrophic factor

DKK3Dickkopf-related protein

DTTDithiothreitol

ECMExtracellular matrix

EGFEpidermal growth factor

G-CSFGranulocyte-colony stimulating factor

GDNFGlial cell derived neurotrophic factor

HGFHepatocyte growth factor

HIF-1Hypoxia inducible factor-1

IGF-1Insulin-like growth factor

IGFBPInsulin-like growth factor-binding protein

ILInterleukin

LS-MSLiquid chromatography-mass spectrometry

MCAMMelanoma cell adhesion molecule

M-CSFMonocyte colony stimulating factor

MEMMinimum essential medium

MIFMacrophage migration inhibitory factor

NG2Neural-glial antigen 2

NGFBeta-nerve growth factor

PDGFPlatelet-derived growth factor

PDGFRβplatelet-derived growth factor receptor beta

PEDFPigment epithelium-derived factor

RNASE4Ribonuclease 4

SCFStem cell factor

SDFStromal cell derived factor

VEGFVascular endothelial growth factor

Electronic supplementary materialThe online version of this article doi:10.1186-s13287-015-0209-8 contains supplementary material, which is available to authorized users.

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Author: Natalia Kalinina - Daria Kharlampieva - Marina Loguinova - Ivan Butenko - Olga Pobeguts - Anastasia Efimenko - Luidmila Age

Source: https://link.springer.com/







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