Proteomic analysis of hypoxia and non-hypoxia secretome mesenchymal stem-like cells from human breastmilk |
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| Authors: | Sri Lilidjanti Widjaja Harsono Salimo Indah Yulianto |
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| Affiliation: | 1. Doctorate Program of Medical Sciences, Faculty of Medicine, Universitas Sebelas Maret, Surakarta, Indonesia;2. Department of Pediatrics, Dr. Moewardi Hospital, Faculty of Medicine, Universitas Sebelas Maret, Surakarta, Indonesia;3. Department of Dermatology and Venereology, Dr. Moewardi Hospital, Faculty of Medicine, Universitas Sebelas Maret, Surakarta, Indonesia;4. Department of Obstetrics and Gynecology, Dr. Moewardi Hospital, Faculty of Medicine, Universitas Sebelas Maret, Surakarta, Indonesia |
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| Abstract: | IntroductionBreastmilk contains proteins and cells which have stem cell properties. The human breastmilk stem cell mimick mesenchymal stem cells and expresses pluripotency genes. The protein level of breastmilk is high in colostrum and gradually subsides in the first year of lactation. The mesenchymal stem cells from breastmilk can be an alternative source of stem cells that can potentially affect cardiovascular therapy. This study aimed to identify the proteomic analysis of secretome mesenchymal stem-like cells under hypoxia compared to non-hypoxia from human breastmilk stem cells.Material and methodsThe human breastmilk was collected from six healthy breastfeeding women and transported to the laboratory under aseptic conditions. The breastmilk cells were isolated then cultured. After 72 h, the human breastmilk stem cells reached confluence then cleaned up and isolated in serum-free media (spheroid) to allow serial passaging every 48 h. The acquisition stem cell was made with flow cytometry. The cells were divided into hBSC secretomes under hypoxia (A) and non-hypoxia (B) and analyzed for LC-MS to identify the peptide structure.ResultsThe human breastmilk cells contained several mesenchymal stem-like cells in density 2.4 × 106 cell/mL for hypoxia and 2 × 106 cell/mL for non-hypoxia conditions. The human breastmilk stem cell surface markers derived from the third cell passage process were 93.77% for CD44, 98.69% for CD73, 88.45% for CD90, and 96.30% for CD105. The protein level of secretome mesenchymal stem -like cells under hypoxia was measured at 5.56 μg/mL and 4.28 μg/mL for non-hypoxia. The liquid chromatography-mass spectrometry analysis identified 130 and 59 peptides from hypoxia and non-hypoxia of the human breastmilk stem cell secretome sequentially. Some important proteomics structures were found in the hypoxic human breastmilk stem cell secretome, such as transforming growth factor-β, VE-cadherin, and caspase.ConclusionThe human breastmilk cells contain mesenchymal stem-like cells and a high concentration of CD44, CD73, CD90, and CD105 as surface markers at third passage culture. The hypoxic hBSC secretome produces a higher protein level compare to non-hypoxia. The transforming growth factor -β was found in the hypoxic hBSC secretome as a modulator of VEGF-mediated angiogenesis. |
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| Keywords: | hBSC Mesenchymal stem-like cell Secretome Hypoxia LC-MS AFP},{#name:keyword,$:{id:k0035},$$:[{#name:text,_:Alpha-Fetoprotein ATP},{#name:keyword,$:{id:k0045},$$:[{#name:text,_:Adenosine Triphosphate BD},{#name:keyword,$:{id:k0055},$$:[{#name:text,_:Becton Dickinson BMPR-II},{#name:keyword,$:{id:k0065},$$:[{#name:text,_:Bone morphogenetic protein type II BSA},{#name:keyword,$:{id:k0075},$$:[{#name:text,_:Bovine Serum Albumin cDNA},{#name:keyword,$:{id:k0085},$$:[{#name:text,_:complementary Deoxyribonucleic Acid EHD3},{#name:keyword,$:{id:k0095},$$:[{#name:text,_:EH Domain-containing Protein 3 FACS},{#name:keyword,$:{id:k0105},$$:[{#name:text,_:Fluorescence-Activated Cell Sorting FBS},{#name:keyword,$:{id:k0115},$$:[{#name:text,_:Fetal Bovine Serum hBSC},{#name:keyword,$:{id:k0125},$$:[{#name:text,_:Human Breastmilk Stem Cell HIF-1α},{#name:keyword,$:{id:k0135},$$:[{#name:text,_:Hypoxia Inducible Factor-1α IGF1},{#name:keyword,$:{id:k0145},$$:[{#name:text,_:Insulin-like Growth Factor 1 LALBA},{#name:keyword,$:{id:k0155},$$:[{#name:text,_:α-Lactalbumin LC-MS},{#name:keyword,$:{id:k0165},$$:[{#name:text,_:Liquid Chromatography-Mass Spectrometry LF},{#name:keyword,$:{id:k0175},$$:[{#name:text,_:Lactoferrin MAPK},{#name:keyword,$:{id:k0185},$$:[{#name:text,_:Mitogen-Activated Protein Kinase MPZL1},{#name:keyword,$:{id:k0195},$$:[{#name:text,_:Myelin Protein Zero-like Protein 1 MPS},{#name:keyword,$:{id:k0205},$$:[{#name:text,_:Multi Proliferative Supplement mRNA},{#name:keyword,$:{id:k0215},$$:[{#name:text,_:messenger Ribonucleic Acid MSC},{#name:keyword,$:{id:k0225},$$:[{#name:text,_:Mesenchymal Stem Cell PBS},{#name:keyword,$:{id:k0235},$$:[{#name:text,_:Phosphate-buffered Saline SDS},{#name:keyword,$:{id:k0245},$$:[{#name:text,_:Sodium Dodecyl Sulfate SMA},{#name:keyword,$:{id:k0255},$$:[{#name:text,_:Smooth Muscle Actin TGF-β},{#name:keyword,$:{id:k0265},$$:[{#name:text,_:Transforming Growth Factor-Beta VEGF},{#name:keyword,$:{id:k0275},$$:[{#name:text,_:Vascular Endothelial Growth Factor SMAD},{#name:keyword,$:{id:k0285},$$:[{#name:text,_:Signals Mothers Against the Decapentaplegic |
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