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- Bartaula-Brevik, Sushma, et al.
(författare)
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Angiogenic and Immunomodulatory Properties of Endothelial and Mesenchymal Stem Cells
- 2016
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Ingår i: Tissue Engineering. Part A. - : Mary Ann Liebert. - 1937-3341 .- 1937-335X. ; 22:3-4, s. 244-252
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Tidskriftsartikel (refereegranskat)abstract
- It has been suggested that the effect of implanted cells on the local environment is important when selecting the appropriate cell type for tissue regeneration. Our aim was to compare the local tissue response to implanted human mesenchymal stem cells (MSC) and human umbilical vein endothelial cells (EC). MSC and EC were cultured in poly(l-lactide-co-1,5-dioxepan-2-one) scaffolds for 1 week in a bioreactor system, after which they were implanted subcutaneously in NOD/SCID mice. After 3 weeks, scaffolds were retrieved, and the mRNA expression of selected genes involved in hypoxia and inflammation was examined by real-time reverse transcription polymerase chain reaction and correlated with immunofluorescent staining for corresponding proteins. The Toll-like receptor signaling pathway was examined by superarray hybridization. The expression of 53 angiogenesis-related proteins was investigated by a proteome profiler angiogenesis antibody array kit. Vascularization was quantified using immunohistochemistry for CD31. The expression of hypoxia-inducible factors and biomarkers for angiogenesis was more strongly upregulated in response to implanted EC than to MSC, suggesting a higher sensitivity to low oxygen tension among EC. Hypoxic signaling was increased after implantation of EC compared with MSC, leading to a prolonged acute inflammatory phase that promoted ingrowth of vascular cells and establishment of the circulation. Inflammatory cytokines were also differently expressed at the gene and protein levels in the two experimental groups, resulting in altered recruitment of acute and chronic inflammatory cells. The end result of these differences was increased vessel formation within the constructs in the EC group.
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| 2. |
- Shanbhag, Siddharth, et al.
(författare)
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Characterization of Human Gingiva- and PDL-derived Progenitor Cells in Xeno-free Conditions
- 2018
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Objectives: Periodontal tissues represent a clinically relevant and less invasive source of progenitor cells compared to bone marrow, for periodontal- and/or alveolar bone-tissue engineering (P-/BTE). The safety and efficacy of clinical-grade stem cells can be enhanced via substitution of xenogeneic supplements and three-dimensional (3-D) cell culture, respectively, to simulate the in vivo microenvironment more closely. The objective of this study was to comprehensively characterize progenitor cells derived from human gingiva (GPCs) and periodontal-ligament (PDL; PPCs) in xeno-free conditions for use in P-/BTE. Methods: In preliminary experiments, pooled human platelet lysate (PL) was identified as the optimal xeno-substitute for fetal bovine serum (FBS) in stem cell cultures. Primary GPCs and PPCs were isolated and expanded as monolayers in PL- and FBS-supplemented media; passage 3-5 cells were used in experiments. Bone marrow mesenchymal stem cells (BMSCs) were used as a reference. Growth kinetics were compared via cell proliferation and colony forming-unit (CFU) assays. GPCs and PPCs were characterized via cytometric expression of stromal markers, multi-lineage (osteogenic, adipogenic and chondrogenic) differentiation potential, and secretory cytokine profiles. 3-D sphere cultures of GPCs and PPCs were established, and the expression of stemness- and osteogenesis-related markers in 3-D and monolayer cultures was evaluated via gene expression and immunocytochemistry. Results: GPCs and PPCs in both FBS and PL showed characteristic fibroblastic morphology and stromal phenotype (highly positive for CD105/CD90/CD73 and negative for CD34/CD45/HLA-DR). Cell proliferation and CFU efficiency were superior in PL compared to FBS. GPCs and PPCs expanded in both PL and FBS showed multi-lineage differentiation comparable to BMSCs; notably, osteogenic differentiation was enhanced in GPCs expanded in PL. 3-D spheres of GPCs and PPCs were formed and maintained for up to 7 days in xeno-free suspension culture. Notably, expression of stemness- (Sox2, Oct4, Nanog) and osteogenesis-related markers (Runx2, Osx, BMP2) was significantly upregulated in GPC- and PPC-derived spheres vs. monolayers. Conclusions: PPCs and particularly GPCs, due to easy access, cultured in 3-D xeno-free conditions represents a promising strategy for P-/BTE. Student Presenter This abstract is based on research that was funded entirely or partially by an outside source: International Team for Implantology (117/2015), Helse Vest Norway (912048) Disclosure Statement: The submitter must disclose the names of the organizations with which any author have a relationship, the nature of the relationship, and the clinical or research area involved. The following is submitted: NONE
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