|
| MSC source | Culture mode | Medium | Results | Ref. |
|
| 3D spheroid cultures |
| Human femoral heads | 3D spheroid culture in spinner flasks and rotating wall vessels | αMEM + 15% FBS | (i) Decrease in surface marker expression levels
(ii) Decreased cell size
(iii) Enhanced osteogenic and adipogenic differentiation
(iv) Differing gene expression profile | [77] |
| Human umbilical cord blood | Spheroids (hanging drop method) | DMEM + 10% FBS, 1% L-glutamine | (i) IL-2Rα, IL-7, IL-16, MCP-3, TGF-β3, and VEGF detected only in spheroid CM
(ii) Significant increase in IL-6, MCP-1, LIF, G-CSF, and SDF-1α
(iii) Decrease in TGF-β1 and TGF-β2 levels
(iv) Decreased chemotactic index of CD14+ cells
(v) Enhanced capability to promote signal factors secretion | [83] |
| Human bone marrow | 3D spheroids (hanging drop method) | CCM + 17% FBS | (i) More effective in suppressing inflammatory responses in the coculture system with LPS-activated macrophages
(ii) Maximally expressed TSG-6
(iii) Expressed high levels of stanniocalcin-1, IL-24, TNF-α-related apoptosis inducing ligand, and CD82
(iv) 1/4 of the volume of monolayer cells | [82] |
| Human bone marrow | 3D spheroids (hanging drop method) | CCM + 17% FBS | (i) Inhibited LPS-stimulated macrophages from secreting proinflammatory cytokines TNF-α, CXCL2, IL-12p40, and IL-23
(ii) Increased secretion of anti-inflammatory cytokines IL-10 and IL-1rα | [28] |
| Human adipose tissue | 3D spheroids in suspension using ultra low attachment plates | αMEM + 10% FBS | (i) Enhanced production of VEGF, SDF, and HGF
(ii) Lowered expression of proapoptotic markers | [78] |
|
| Oxygen tension (hypoxia/anoxia) |
| Human adipose tissue | Hypoxia (1% O2) | DMEM | (i) Higher HIF-1α expression
(ii) Increased release of EVs
(iii) Induced overexpression of miRNAs implicated in inflammatory (miR-223, -146b), proliferative, and differentiative phases (miR-126, -199a) of the healing process
(iv) Enhanced muscle regeneration process | [89] |
| Human | Anoxia (0.1% O2), hypoxia (5% O2) | αMEM +5 g/L glucose | (i) CM from anoxic conditions enhanced chemotactic and proangiogenic properties and reduced inflammatory mediator content
(ii) Enhanced expression of VEGF-A, VEGF-C, IL-8, RANTES, and monocyte chemoattractant protein 1 | [86] |
| Human adipose tissue | Hypoxia (5% O2) | RKCM | (i) Promoted antiapoptotic effects
(ii) Higher levels of GM-CSF, VEGF, IL-6, and IGF-1
(iii) Lower levels of TGF-β1 | [92] |
| Human umbilical cord Wharton jelly | Hypoxia (5% O2) | PPRF-msc6 | (i) Increased secretion profile
(ii) Upregulated thymosin-beta and EF-2 significantly
(iii) Enhanced neuroregulatory secretome profile | [90] |
| Human bone marrow | Hypoxia (1% O2) | DMEM + 10% FBS + 2 mM L-glutamine | (i) Upregulated protein level of vimentin, fibronectin, and N-cadherin
(ii) Enhanced stemness genes Oct4, Nanog, Sall4, and Klf4
(iii) Higher levels of osteocalcin and osteopontin
(iv) Reduced levels of COL2A1, COMP, and aggrecan
(v) Lower expression of adipsin, FASN, and FABP4
(vi) Upregulated IGFs, VEGF, EGF, GCSF, GM-CSF, TGF-β1, and TGF-β2 | [93] |
| Human bone marrow | Hypoxia (1% O2) with serum starvation | Opti-MEM + 1% L-glutamine | (i) Significant increases in rate-limiting proteins of glycolysis and the NRF2/glutathione pathway
(ii) Upregulated angiogenic associated pathways of PDGF, EGF, and FGF
(iii) Microvesicle secretion decreased, exosome secretion substantially increased | [88] |
| Murine bone marrow | Repeated cycles of anoxia | StemPro MSC SFM | (i) miR-11, miR-22, miR-24, miR-199a-3p, and miR-210 upregulated in exosomes | [91] |
| Human bone marrow | Hypoxia for 30, 60, or 90 min | Unknown | (i) The 60 min group had the greatest protective effect on endotoxin-induced acute lung injury model | [94] |
|
| Mechanical stimuli |
| Human bone marrow | TGF-β1 stimulation (1 ng/mL) or mechanical load (multiaxial shear and compression) in fibrin-poly(ester-urethane) scaffolds | αMEM + 10% FBS + 5 ng/mL bFGF | (i) TGF-β1 stimulation and load had distinct effects, both enhanced chondrogenic profile compared to control
(ii) Nitrite content in media higher in loaded groups
(iii) TGF-β1 enhanced expression of leptin, leptin receptor, and MDC
(iv) Load enhanced expression of uPAR, LAP, MIP3α, angiogenin, ALCAM, angiopoietin 2, osteoprotegerin, and DR6; reduced expression of GRO
(v) Both TGF-β1 and load enhanced the expression of BLC, MCP3, MIF, VEGF, MMP13, and PDGFaa | [100] |
| Human bone marrow | Computer-controlled bioreactors, on Cytodex 3 microcarriers (2 g/L) | PPRF-msc6 | (i) Enhanced the neuroregulatory profile of secretome
(ii) Increased the secretion of Cys C, GDN, Gal-1, and PEDF
(iii) Upregulation of miR-16
(iv) Number of CNS regulators only detected in CM of bioreactor cultured MSCs
(v) Upregulation of classical trophic factors BDNF, VEGF, and IGF-1 | [16] |
| Human bone marrow | Bioreactors | DMEM + 10% FBS | (i) Enhanced angiogenesis by CM from mechanically stimulated MSCs via FGFR and VEGFR signalling cascades
(ii) Enrichment of MMP-2, TGF-β1, and bFGF | [102] |
| Human | PAM hydrogels of various rigidity | DMEM-low glucose + 10% FBS | (i) VEGF, angiogenin, and IGF upregulated with increasing elastic modulus
(ii) EGF, IL-6, and IL-8 were not stiffness-dependent | [103] |
| Adipose tissue | Fibrous scaffolds of variously aligned fibers | α-MEM + 10% FBS | (i) Higher levels of anti-inflammatory and proangiogenic cytokines were produced from cells seeded on electrospun scaffolds
(ii) CM from scaffold cultures accelerated wound closure and macrophage recruitment in wound bed | [163] |
|
| Electromagnetic stimuli |
| Equine adipose tissue | Static magnetic field (0.5 T) | DMEM/F12 + 10% FBS | (i) Reached doubling time earlier, colony-forming potential higher
(ii) Considerable increase in the number of secreted microvesicles
(iii) Release of BMP-2, VEGF, and p53 increased
(iv) Reduced release of TNF-α | [113] |
|
| Biochemical stimuli |
| Human bone marrow | IFN-γ and TNF-α stimulation | DMEM-low glucose | (i) Elevated secretion levels of IL-6, HGF, VEGF, and TGF-β | [119] |
| Murine bone marrow | IFN-γ and either TNF-α, IL-1α, or IL-1β stimulation | α-MEM+ 10% FBS + 2 mM glutamine | (i) Provoked the expression of CXCL-9 and CXCL-10 and inducible nitric oxide synthase | [118] |
| Human adipose tissue | TGF-β1 stimulation | DMEM + 0.1% BSA + 1% glutamine | (i) Upregulated secretion of PIGF, IGFBP-3, LIF, OSM, IL-4, IL-7, IL-13, CXCL9, CCL26, and OPN
(ii) Downregulated secretion of CCL7, CCL11, CXCL6, OPG, IL-5, IL-10, CCL8, CXCL1, CXCL10, HGF, leptin, FGF-7, and GM-CSF | [123] |
| Adipose tissue | TNF-α stimulation | MesenPRO RS Basal Medium + 2 mM L-glutamine + MesenPRO RS Growth Supplement | (i) TNF-α-preconditioned ASCs secreted exosomes with elevated Wnt-3a content
(ii) Enhanced proliferation and osteogenic differentiation in human primary osteoblastic cells | [120] |
| Human umbilical cord | LPS preconditioning | DMEM-low glucose + 10% FBS and sigma serum-free medium | (i) Improved regulatory abilities for macrophage polarization and resolution of chronic inflammation
(ii) Unique expression of miR-let7b | [115] |
| Human adipose tissue | LPS preconditioning | DMEM-low glucose | (i) Enhanced mRNA expression of IL-6, TNF-α, HGF, and VEGF
(ii) Enhanced liver regeneration in partially hepatectomized mice | [116] |
| Human adipose tissue | H2O2 stimulation | α-MEM + 10% exo-free FBS | (i) Exosomes that had been H2O2-stimulated enhanced skin flap recovery and capillary density | [121] |
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