Departmental Papers (Dental)

Document Type

Journal Article

Date of this Version

10-2018

Publication Source

Molecular Medicine Reports

Volume

18

Issue

4

Start Page

3555

Last Page

3562

DOI

10.3892/mmr.2018.9410

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.

Comments

At the time of publication, author Chenshuang Li was affiliated with the Peking University School of Stomatology. Currently, (s)he is a faculty member at the School of Dental Medicine at the University of Pennsylvania.

Keywords

CONTACTIN-ASSOCIATED PROTEIN-LIKE 4 (CNTNAP4), 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

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Date Posted: 10 February 2023