Isolation and Characterization of Minipig Perivascular Stem Cells for Bone Tissue Engineering
Penn collection
Degree type
Discipline
Subject
LIGAND/RECEPTOR-LIKE INTERACTION
NELL-1/CNTNAP4 AXIS
NEURAL EGFL-LIKE 1 (NELL-1)
OSTEOGENESIS
Amino Acid Sequence
Animals
Animals
Newborn
Bacteriophage T7
Bone Marrow
Calcium-Binding Proteins
Cell Line
Cell Lineage
Cell Membrane
Gene Deletion
Glycoproteins
Humans
Integrases
Membrane Proteins
Mice
Inbred C57BL
Mice
Knockout
Models
Biological
Nerve Tissue Proteins
Osteogenesis
Protein Binding
Protein Domains
Signal Transduction
Skull
alkaline phosphatase
bacteriophage DNA
bone morphogenetic protein 2
bone sialoprotein
cell surface receptor
collagen type 1 alpha 1
collagen type 1 alpha 2
contactin
contactin associated protein like 4
membrane receptor
mitogen activated protein kinase
neural egfl like 1
neurexin
osteocalcin
osteopontin
unclassified drug
Wnt1 protein
calcium binding protein
Cntnap4 protein
mouse
cre recombinase
glycoprotein
integrase
membrane protein
Nell1 Cluster of differentiation 146
Cluster of differentiation 34
Flow cytometry
Osteogenic potential
Perivascular stem cells
Adipogenesis
Animals
Antigens
CD34
CD146 Antigen
Cell Separation
Flow Cytometry
Humans
Leukocyte Common Antigens
Male
Mesenchymal Stem Cells
Osteogenesis
Stem Cells
Swine
Swine
Miniature
Tissue Engineering
alizarin red s
CD146 antigen
CD34 antigen
cell surface marker
collagen type 1
collagen type 1 alpha 1
eosin
hematoxylin
messenger RNA
osteocalcin
peroxisome proliferator activated receptor gamma
receptor type tyrosine protein phosphatase C
unclassified drug
CD146 antigen
CD34 antigen
receptor type tyrosine protein phosphatase C
adipogenesis
adventitia
animal cell
animal tissue
Article
bone development
bone tissue
cell differentiation
cell isolation
cell population
cell structure
colony forming unit
controlled study
fibroblast
fluorescence activated cell sorting
human
human cell
human cell culture
in vitro study
liposuction
male
minipig
mRNA expression level
nape
nonhuman
pericyte
perivascular stem cell
quantitative analysis
reverse transcription polymerase chain reaction
staining
stem cell
subcutaneous fat
tissue engineering
tissue structure
white adipose tissue
animal
cell separation
cytology
flow cytometry
mesenchymal stem cell
minipig
pig
procedures
stem cell
tissue engineering
Dentistry
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Grant number
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Abstract
Human subcutaneous adipose tissue has been recognized as a rich source of tissue resident mesenchymal stem/stromal cells (MSC) in recent years. The current study was designed to sort the minipig (mp) perivascular stem cells (PSCs) and investigate the osteogenic potential. Purification of human PSCs was achieved via fluorescence-activated cell sorting (FACS) from human liposuction samples [cluster of differentiation (CD)45-CD34-CD146+ perithelial cells and CD45-CD34+CD146- adventitial cells]. Subsequently, PSCs were isolated from mp adipose tissue samples (n=9), characterized and, using purified mpPSCs (obtained by FACS, which is used in human PSC purification), the mpPSC osteogenic and adipogenic potential was evaluated by Alizarin Red S and Oil Red O staining in vitro, respectively. The cell morphometry was observed following cell isolation and culture, and hematoxylin and eosin staining was performed to identify the fat tissue structure and vascular distribution. Osteogenic and adipogenic differentiation-associated gene expression levels were analyzed by reverse transcription-quantitative polymerase chain reaction. The results demonstrated that the same antigens used for human PSC identification and isolation were working in mp tissue (CD45, CD146 and CD34). The two cell groups: CD45-CD34-CD146+ pericytes and CD45-CD34+CD146- adventitial cells were successfully isolated from the subcutaneous fat in the posterior neck of mps, mpPSCs accounted for 8.6% of the stromal vascular fraction (SVF) with 1.4% pericytes and 7.2% adventitial cells. mpPSCs demonstrated characteristics of MSCs, including cell surface marker expression, colony forming unit-fibroblast inclusion, and the stronger osteogenic and adipogenic differentiation potential than that of the non-selected vascular stromal cells. The mRNA expression levels of osteocalcin, collagen, type I, α1 and peroxisome proliferator-activated receptor-γ in the mpPSCs group were significantly higher than those of the unsorted pSVF group (P<0.05). Thus, the current study successfully isolated and cultured CD146+ and CD34+ cell populations from mp tissues, characterized the cells' PSC-like phenotype and identified their distinctly osteogenic and adipogenic potential. © Spandidos Publications. All rights reserved.