12 research outputs found
Human amnion-derived mesenchymal stem cells attenuate acute lung injury in two different acute lung injury mice models
Acute lung injury (ALI) is one of the most common clinical emergencies with limited effective pharmaceutical treatment in the clinic, especially when it progresses to acute respiratory distress syndrome (ARDS). Currently, mesenchymal stem cells (MSCs) exhibit specific superiority for ALI/ARDS treatment. However, stem cells from different sources may result in controversial effects on similar disease conditions. This study aimed to determine the effects of human amnion-derived mesenchymal stem cells (hAMSCs) on two different ALI mice model. The administered hAMSCs effectively accumulated in the lung tissues in all hAMSC-treated groups. Compared with the model and 1% human serum albumin (HSA) groups, high-dose hAMSCs (1.0 × 106 cells) group significantly alleviated alveolar-capillary permeability, oxidative stress, inflammatory factors level and histopathological damage. In addition, the NF-κB signaling pathway is one of the key pathways activated during lipopolysaccharide (LPS) or paraquat (PQ)-induced lung injury. Our results indicated that hAMSCs (1.0 × 106 cells) obviously inhibited the expression of p-IKKα/β, p-IκBα, and p-p65 in the lung tissue (p < 0.05). The high-dose (HD) hAMSC treatment exerted beneficial therapeutic effects on ALI mice models without detectable adverse reactions. The therapeutic effect of hAMSCs might involve NF-κB signaling pathway inhibition. hAMSC treatment is a potential candidate therapy for ALI
hESCs growing eiMEFs expressed stem cell markers.
<p><b>A,</b> Immunostaining showed pluripotent markers of NANOG, OCT4, SSEA4 and TRA-1-81 expressed in hESCs cultured on eiMEFs. Scar bar, 100 μm. <b>B,</b> FACS assay showed that during consecutive incubation (day 7 (passage 46), day 14 (passage 47), day 21 (passage 48) and day 28 (passage 49), high percentage of cells positive for OCT4 (98.3%±2.55%, 94.3%±2.20%, 96.2%±2.55%, 97.0%±2.95%), NANOG (97.5%±2.15%, 94.4%±1.65%, 95.5%±1.98%, 98.0%±1.95%), SSEA4 (, 97.9%±2.40%, 93.4%±2.10%, 97.4%±2.54%, 97.4%±2.36%) and TRA-1-81 (98.6%±3.12%, 96.1%±1.85%, 96.3%±2.35%, 97.6%±2.88%) stablely expressed in hESCs cultured eiMEFs.</p
hESCs cocultured with eiMEFs were pluripotency <i>in vivo</i> and <i>in vitro</i>.
<p><b>A</b>, Teratomas formed after hESCs were injected into nud mice for 8 weeks. <b>B</b>, Gland tissure in the hESCs-derived teratoma. Scar bar, 50 μm. <b>C</b>, Adipose tissue and muscles in the hESCs-derived teratoma. Scar bar, 50 μm. <b>D</b>, Epidermal and neural tissues in the hESCs-derived teratoma. Scar bar, 50 μm. <b>E</b>, hESCs formed embryoid bodies (hEBs). <b>F</b>, Hoechst staining showed cell nucleus (blue). Scar bar, 10 μm. <b>G</b>, hESCs-derived immature neurons were indicated by BIII-tubulin (TUJ1, green). <b>H</b>, hESCs-derived dopaminergic neuron cells were marked by tyrosine hydroxylase (TH, red). <b>I</b>, Picture merged from F, G and H. <b>J</b>, Quantitative PCR (qPCR) analysis on the expression level of gene markers of the three-germ layers. <i>NESTIN</i> and <i>SOX1</i> (ectoderm markers), <i>T</i> and <i>CD31</i> (mesoderm markers), <i>AFP</i> and <i>SOX17</i> represented (entoderm markers). There were no statistical differences between the expression level of marker genes in eiMEFs-hEBs and miMEFs-hEBs. miMEFs-hEBs, embryoid bodies derived from hESCs cultured on miMEFs. eiMEFs-hEBs, embryoid bodies derived from hESCs cultured on eiMEFs.</p
No residual ethanol detected in old culture media harvested from eiMEF culture.
<p>Old culture media was harvested at day 1, 2, 3, 4, 5, 6 and 7 after MEFs plated. The concentrations of residual ethanol in harvested media were measured by ethanol detection kit. Negligible, background levels of ethanol were detected in both eiMEFs group and untreated MEFs group. And Statistical analysis showed there was no significant difference between the two groups.</p
Designations, sequences, and the sizes of real-time PCR amplicons.
<p>Designations, sequences, and the sizes of real-time PCR amplicons.</p
10% ethanol-treated MEFs supported hESCs growth.
<p><b>A</b>, Alkaline phosphatase (AKP) staining analysis on hESCs cultured on MEFs treated by different concentrations of ethanol (5%, 10%, 20% and 30%). <b>B</b>, The morphology of hESCs cultured on MEFs treated by different concentrations of ethanol (5%, 10%, 20% and 30%) during growth day 1, 3 and 5. <b>C,</b> AKP staining analysis on hESCs cultured on ethanol-treated MEFs plated with different densities. <b>D</b>, The morphology of hESCs cultured on frozen-thawed eiMEFs. MC, mitomycin C. ET, ethanol. miMEFs, 10 ug/ml mitomycin C inactivated mouse embryonic fibroblasts. eiMEFs, 10% ethanol inactivated mouse embryonic fibroblasts. Red arrows indicate differentiated cells. Scar bar, 100 μm.</p
Therapeutic effects and mechanism of human amnion-derived mesenchymal stem cells on hypercoagulability in a uremic calciphylaxis patient
AbstractCalciphylaxis is a rare cutaneous vascular disease that manifests with intolerable pains, non-healing skin wounds, histologically characterized by calcification, fibrointimal hyperplasia, and microvessel thrombosis. Currently, there are no standardized guidelines for this disease. Recent studies have recognized a high prevalence of thrombophilias and hypercoagulable conditions in calciphylaxis patients. Here, we report a case of uremic calciphylaxis patient whom was refractory to conventional treatments and then received a salvage strategy with intravenous and local hAMSC application. In order to investigate the therapeutic mechanism of hAMSCs from the novel perspective of hypercoagulability, coagulation-related indicators, wound status, quality of life and skin biopsy were followed up. Polymerase chain reaction (PCR) was performed to determine the distribution of hAMSCs in multiple tissues including lung, kidney and muscle after infusion of hAMSCs for 24 h, 1 week and 1 month in mice aiming to investigate whether hAMSCs retain locally active roles after intravenous administration. Improvement of hypercoagulable condition involving correction of platelet, D-dimer and plasminogen levels, skin regeneration and pain alleviation were revealed after hAMSC administration over one-year period. Skin biopsy pathology suggested regenerative tissues after 1 month hAMSC application and full epidermal regeneration after 20 months hAMSC treatment. PCR analysis indicated that hAMSCs were homing in lung, kidney and muscle tissues of mice even until tail vein injection of hAMSCs for 1 month. We propose that hypercoagulability is a promising therapeutic target of calciphylaxis patients, which can be effectively improved by hAMSC treatment