ABSTRACT
BACKGROUND: Perivascular cells have been shown to be the precursor cells of mesenchymal stem cells, which regulate the behavior of hematopoietic stem cells and support hematopoiesis through cell-to-cell contact or paracrine effects. Hematopoietic support of human skeletal muscle-derived pericytes/perivascular cells (hMD-PCs) remains to be studied. OBJECTIVE: To identify the biological characteristics of hMD-PCs isolated from human skeletal muscle and to study their supporting effect on umbilical cord blood CD34+ cells in vitro. METHODS: (1) hMD-PCs with phenotype CD146+ CD56-CD34-CD144-CD45- were sorted from human skeletal muscle by enzymatic digestion and multiparameter fluorescence-activated cell sorting, and their biological characteristics were identified. (2) The in vitro culture system of umbilical cord blood CD34+ cells co-cultured with human CD146+ hMD-PCs (experimental group) or with human bone marrow mesenchymal stem cells (positive control group) was established. After 1, 2 and 4 weeks of co-culture, the number of cells, the colony formation ability and immunophenotype were measured and statistically analyzed. RESULTS AND CONCLUSION: (1) CD146+ hMD-PCs were sorted by multiparameter fluorescence-activated cell sorting and the purity was (91.5±1.85)% (n=5). CD146+ hMD-PCs expressed mesenchymal surface markers CD73, CD90, CD105, CD44, and did not express hematopoietic cell and endothelial cell markers CD45, CD34, and CD31. After induced culture, CD146+ hMD-PCs could differentiate into osteoblasts, chondrogenesis, adipocytes and myoblasts. (2) There were no significant differences in the cell number, colony f ormation ability or immunophenotype (CD45+, CD34+ CD33-, CD14+, CD10+/CD19+) between experimental and positive control groups (P > 0.05, n=6). The number of cells in the blank control group without feeder was significantly decreased at 1 week of culture, and there was almost no cell survival at 2 weeks of culture. (3) In summary, CD146+ hMD-PCs, like human bone marrow mesenchymal stem cells, have hematopoietic support capacity in vitro.
ABSTRACT
BACKGROUND: Pericytes and other perivascular stem cells are gaining increasing attention in bone tissue engineering. Pericytes have been long thought to regulate blood pressure and promote angiogenesis. However, it is now considered to have the characteristics of mesenchymal stem cells, including pluripotency, self-renewal, immune regulation, and tissue repair, showing strong regenerative potential in common orthopedic diseases such as fractures, nonunion, vertebral fusion, ligament rupture, and cartilage injury. OBJECTIVE: To summarize the related applications of pericytes in orthopedics, and to explore mechanisms, strengths and limitations.METHODS: With “pericytes, perivascular stem cells, repair, therapy, orthopedics” as Chinese and English retrieval terms, we systematically searched databases including PubMed, Web of Science, CNKI and WanFang, from inception to August 2019 for the articles concerning the treatment of orthopedic diseases with pericytes. A total of 112 related documents were retrieved. After reading the full text, 55 eligible documents were selected. RESULTS AND CONCLUSION: Pericytes are a kind of emerging stem cells that have been reported in many studies regarding the treatment of orthopedic diseases, and have a greatly therapeutic prospect. There are abundant pericytes in the body with no need for in vitro culture. The repair mechanism of pericytes is mainly related to paracrine mediation, with self-differentiation as a secondary mechanism. However, its specific mechanism remains unclear, which needs further investigations.
ABSTRACT
Objective:To study the effect of Shenshuai Xiezhuo decoction and its deficiency tonifying and pathogen eliminating components on renal interstitial fibrosis in UUO rats. Method:A rat model of unilateral ureteral obstruction (UUO) was established through ligation of a unilateral ureter. The rats were divided into six groups: sham operation group, model group, benazepril group, Shenshuai Xiezhuo decoction group, Buxufang group, and phlegm group, with 24 rats in each group. On the third day after operation, the rats in the Shenshuai Xiezhuo decoction group, Buxufang group, and phlegm group were given Shenshuai Xiezhuo decoction concentrating agent at a dose of 8.0 g·kg-1·d-1, the rats in the benazepril group were given benazepril 1.5 mg·kg-1·d-1, and the rats in the sham operation group and the model group were given the same volume of saline. On the 7th, 14th and 21st days after operation, the expressions of peripheral cells and relevant signal pathway markers in renal tissue were detected by Western blot and Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) respectively. Result:The stromal damage score and the interstitial collagen accumulation on the 14th and 21st days after UUO were significantly lower in the Shenshuai Xiezhuo decoction group, Buxu prescription group and Qixie prescription group than those in the model group (P<0.05). Except for the sham operation group, the protein expressions of α smooth muscle actin (α-SMA), platelet derived growth factor A (PDGFA), nerve/glial type 2 chondroitin sulfate glycoprotein (NG-2), vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 1 (VEGFR1), platelet-derived growth factor -β (PDGF-β) and platelet-derived growth factor receptor-β (PDGFR-β) in the kidney tissues of the 14th and 21st days were significantly increased compared with those on the 7th day (P<0.05). On the 21st day after surgery, the expressions of NG-2, VEGFA, VEGFR1 and PDGFR-β in renal tissue of Shenshuai Xiezhuo decoction group were significantly lower than those in the model group (P<0.05). Compared with the model group, relevant expressions of α-SMA, PDGFA, NG-2, VEGFA, VEGFR1, PDGF-β, and PDGFR-β in kidney tissue of Shenshuai Xiezhuo decoction group decreased significantly on the 21st day after operation (P<0.05), and relative expressions of α-SMA, NG-2, VEGFA, VEGFR1 and PDGFR-β in the Shenshuai Xiezhuo decoction group were significantly lower than those in the benazepril group on the 21st day after surgery (P<0.05). Conclusion:Shenshuai Xiezhuo decoction and its deficiency tonifying and pathogen eliminating components have an antagonistic effect on renal interstitial fibrosis in UUO rats. Its mechanism is related to the inhibition of activation of peripheral cells and relevant cell signaling pathways.
ABSTRACT
Understanding the crosstalk mechanisms between perivascular cells (PVCs) and cancer cells might be beneficial in preventing cancer development and metastasis. In this study, we investigated the paracrine influence of PVCs derived from human umbilical cords on the proliferation of lung adenocarcinoma epithelial cells (A549) and erythroleukemia cells (TF-1α and K562) in vitro using Transwell® co-culture systems. PVCs promoted the proliferation of A549 cells without inducing morphological changes, but had no effect on the proliferation of TF-1α and K562 cells. To identify the factors secreted from PVCs, conditioned media harvested from PVC cultures were analyzed by antibody arrays. We identified a set of cytokines, including persephin (PSPN), a neurotrophic factor, and a key regulator of oral squamous cell carcinoma progression. Supplementation with PSPN significantly increased the proliferation of A549 cells. These results suggested that PVCs produced a differential effect on the proliferation of cancer cells in a cell-type dependent manner. Further, secretome analyses of PVCs and the elucidation of the molecular mechanisms could facilitate the discovery of therapeutic target(s) for lung cancer.