Proteomic analysis of hypoxia and non-hypoxia secretome mesenchymal stem-like cells from human breastmilk

Autor: Sri Lilidjanti Widjaja, Soetrisno, Indah Yulianto, Harsono Salimo
Jazyk: angličtina
Rok vydání: 2021
Předmět:
0106 biological sciences
0301 basic medicine
Angiogenesis
Cell
TGF-β
Transforming Growth Factor-Beta
01 natural sciences
HIF-1α
Hypoxia Inducible Factor-1α
ATP
Adenosine Triphosphate
Biology (General)
Hypoxia
Secretome
PBS
Phosphate-buffered Saline
biology
medicine.diagnostic_test
VEGF
Vascular Endothelial Growth Factor
IGF1
Insulin-like Growth Factor 1
LF
Lactoferrin
SMAD
Signals Mothers Against the Decapentaplegic
LC-MS
Liquid Chromatography-Mass Spectrometry
MSC
Mesenchymal Stem Cell
Cell biology
hBSC
medicine.anatomical_structure
Original Article
Stem cell
General Agricultural and Biological Sciences
MPS
Multi Proliferative Supplement
Mesenchymal stem-like cell
BMPR-II
Bone morphogenetic protein type II
FACS
Fluorescence-Activated Cell Sorting
EHD3
EH Domain-containing Protein 3
QH301-705.5
SMA
Smooth Muscle Actin
cDNA
complementary Deoxyribonucleic Acid
hBSC
Human Breastmilk Stem Cell
Flow cytometry
AFP
Alpha-Fetoprotein
03 medical and health sciences
mRNA
messenger Ribonucleic Acid
medicine
CD90
BD
Becton Dickinson
CD44
Mesenchymal stem cell
MAPK
Mitogen-Activated Protein Kinase
LALBA
α-Lactalbumin
LC-MS
030104 developmental biology
Cell culture
biology.protein
MPZL1
Myelin Protein Zero-like Protein 1
BSA
Bovine Serum Albumin
SDS
Sodium Dodecyl Sulfate
010606 plant biology & botany
FBS
Fetal Bovine Serum
Zdroj: Saudi Journal of Biological Sciences, Vol 28, Iss 8, Pp 4399-4407 (2021)
Saudi Journal of Biological Sciences
Popis: Introduction Breastmilk 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 methods The 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. Results The 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. Conclusion The 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.
Databáze: OpenAIRE
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