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Copyright ©The Author(s) 2020.
World J Stem Cells. Nov 26, 2020; 12(11): 1341-1353
Published online Nov 26, 2020. doi: 10.4252/wjsc.v12.i11.1341
Table 1 Source and isolation methods of mesenchymal stromal cells
Source
Isolation method
Ref.
Bone marrowAspirates cultured and media changed every 3-4 d to select for MSCs[34]
Aspirates layered over Ficoll-Paque density-gradient and plates in tissue culture dish. Adherent cells maintained with periodic passaging[82]
Bone marrow mononuclear cells seeded from single colony-forming unit fibroblasts and selected for by CD105(+)/CD45(-)[83]
Sort bone mononuclear cells based on aldehyde deydrogenase expression (ALDHhighCD45-)[84]
Sort based on CD45−/lowCD271+ phenotype following a microbead-based pre-enrichement[85]
Layer bone marrow over hyluronic acid followed by centrifugation, collect most superficial layer containing the mononuclear cells[86]
Compact boneTrabecular bone fragments rinsed and placed in complete α-MEM/Ham’s F12, confluent monolayers were obtained within 10-20 d[87-89]
Bone cell cultures established by treating bone fragments with collagenase in low Ca2+ medium [9,90,30]
Compact bone fragments obtained, cultured, and isolated. CB-MSCs then undergo trypsinization to reveal enhanced osteogenic capacity[43]
Adipose Place 10-20 mL of washed adipose tissue in 100 mm Petri dish; dissect out yellow tissue; mince tissue finely and place in enzymatic digestion solution; centrifuge and collect pellet for wash; resuspend in complete culture medium[14,13]
Wash lipoaspirate with PBS; enzymatically digest using collagenase 1A solution; spin down cells, wash and plate in complete medium[12]
Bone marrowAspirates cultured and media changed every 3-4 d to select for MSCs[34]
Aspirates layered over Ficoll-Paque density-gradient and plates in tissue culture dish. Adherent cells maintained with periodic passaging[82]
Bone marrow mononuclear cells seeded from single colony-forming unit fibroblasts and selected for by CD105(+)/CD45(-)[83]
Sort bone mononuclear cells based on aldehyde deydrogenase expression (ALDHhighCD45-)[84]
Sort based on CD45−/lowCD271+ phenotype following a microbead-based pre-enrichement[85]
Table 2 Potential clinical scenarios for use of mesenchymal stromal cell therapy
Ref.
Animal model/methods
Findings
Conclusion
In vitro/in vivo
Ogulur et al[91]mCB-MSCs isolated from 6–8 wk old BALB/c micemCB-MSCs significantly reduced cellular immune infiltration and presence of goblet cells as well as the thickness of epithelium, smooth muscle layers, and basement membrane in ovalbumin induced chronic asthmatic miceInflammation in distal and proximal airways of ovalbumin induced asthmatic mice can be suppressed by use of IV mCB-MSCsIn vivo
Qiao et al[76] CB-MSCs isolated from C57BL/6 mice administered to 8–10 wk old BALB/c miceBALB/c mice exposed to 8 Gy TBI and treated with CB-MSCs showed improved survival, body weight, and CFU-GM counts of bone marrow cells coupled with suppressed Th1 immunity with increased Treg percentages and decreased IFN-γ, CXCR3 and CCR5CB-MSC transplantation post total body irradiation attenuates radiation-induced hematopoietic toxicity and provides immunoprotectionIn vivo
Duran et al[92]Cortical bone–derived stem cells from 12-wk-old EGFP+ transgenic miceImproved 6 wk survival post MI procedure (50.4% to 76.5%) from saline to CB-MSC therapy. Increased expression of proangiogenic paracrine factors (bFGF and VEGF) and differentiation into infarct zoneTreatment with CB-MSCs post MI leads to enhanced survival, cardiac function, and remodelingIn vivo
Cheng et al[93]MSCs isolated from compact bone of Tg26 HIV-1 transgenic miceTransplanted Tg26 HIV-1 MSCs were less effective in protecting renal tubular cells compared to healthy mice MSCs in a cisplatin-induced AKI model due to inferior proliferation and decrease in secretion of protective cytokinesCompact bone MSCs infected with HIV-1 had impaired proliferation, differentiation, and function resulting in less therapeutic potentialIn vivo
Yamachika et al[94]MSCs from compact bone of 5-week-old C57-GFP male miceCells cultured in bFGF-conditioned medium demonstrated trilineage differentiation potential even at passage 24 in contrast to leukemia inhibitory factor-conditioned medium Compact bone MSCs cultured in bFGF-conditioned medium demonstrated bone formation ability in vivoIn vivo
Bakker et al[95]Tibial reaming debris from adult female sheepTreatment with reaming debris, similar to iliac crest, revealed larger callus volume with decreased cartilage in the fracture gap, increased bone volume, and improved toughness at 3 wk with greater torsional stiffness at 6 wkReaming debris has characteristics similar to iliac crest bone that allow it to be an excellent replacement for enhancing healing of bone defects fixed with an intramedullary nailIn vivo
Guo et al[24] Murine mesenchymal progenitor cells (muMPCs) isolated from 2-3 wk old C57BL/6 female mice tibia/femur compact boneCollagenase-digested bone fragments produced muMPCs that inhibited Con A-stimulated splenocyte proliferation and suppressed lymphocyte activation by allogeneic cellular stimuli in vitro. In addition, muMPCs improved survival of allogeneic skin grafts in vivoUsing this protocol allows acquiring of muMPCs with similar properties to marrow counterparts, which allows them to be used in future investigations with mouse modelsIn vivo
Lim et al[96]hABMSCshABMSCs exposed to low-intensity pulsed ultrasound revealed increased ALP, expression levels of CD29, CD44, COL1, and OCN, and calcium depositionTreatment with LIPUS could improve the cell viability and osteogenic differentiation of hABMSCsIn vitro
Lim et al[97]hABMSCshABSMSCs treated with extremely low frequency pulsed electromagnetic fields (ELF-PEMFs) revealed 15% increased proliferation at day 5, increased ALP, vinculin, vimentin, and CaM expressions, and enhanced mineralization during osteogenesisExposing hABMSCs with ELF-PEMFs could improve and accelerate the process of early cell proliferation mediated osteogenesisIn vitro
Lim et al[98]hABMSCs harvested from human mandibular alveolar bonehABSMSCs exposed to LFDSS for 10–60 min/d demonstrated improved viability, proliferation, and mineralization in culture with osteoblasts. ALP activity and gene expression of IBSP, COL-I, OCN, and OPN increasedProper intensity and exposure time of LFDSS to hABMSCs can improve their differentiation and maturationIn vitro
Soleimani et al[35]MSCs isolated from 6-8 wk old BALB/c mouse tibial and femoral bone marrowThe protocol states MSCs should be cultured in Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) in a 37 °C–5% CO2 incubator with passage at 2 wk of cultureThis protocol allows development of a purified population of MSCs 3 wk after the initiation of cultureIn vitro
Dominici et al[44]Human multipotent MSCIn standard culture, MSC must be plastic-adherent, express CD105, CD73 and CD90, and lack expression of CD45, CD34, CD14 or CD11b, CD79alpha or CD19 and HLA-DR and demonstrate tridifferentiation in vitroStandard criteria for MSC characterization, will allow for exchange of more uniform data between researchersIn vitro
Wenisch et al[99]Mesenchymal stem cells harvested from HRD of 12 adult patients with closed diaphyseal femoral fracturesWith neuronal induction, MSCs assumed neuronal morphologies and expressed neuron-specific enolase, beta-III-tubulin, neurofilament-H and HNK-1. Similar to immature neurons, MSCs had features of neuritogenesis and synaptogenesis and lacked electrical signalingNeuronal induction allowed initiation of the early neuronal differentiation, but exposure to non-neurological stressors led to necrotic alterationsIn vitro
Wenisch et al[100]Mesenchymal stem cells harvested from HRD of 12 adult patients with closed diaphyseal femoral fracturesAfter multiple passages, HRD-derived cells and MSCs maintained a nondifferentiated phenotype and showed osteogenic and neuronal pathway differentiation ability after inductionHuman reaming debris provides a multipotent stem cells which have the ability to grow and proliferate in vitroIn vitro
Tuli et al[90]Collagenase-treated human trabecular bone chipsCollagenase-treated trabecular bone fragments contain cells that stain positive for CD73, STRO-1, and CD105, and negative for CD34, CD45, and CD144 with tridifferentiation potentialTrabecular bone-derived cells maintain a nondifferentiated phenotype and display tridifferentiation potential with long-term in vitro cultureIn vitro