Journal: Orphanet Journal of Rare Diseases

Article Title: Impaired osteoblast and osteoclast function characterize the osteoporosis of Snyder - Robinson syndrome

doi: 10.1186/s13023-015-0235-8

Figure Lengend Snippet: Characterization of the bone and osteoblast pathology. A . Photograph of the bone biopsy. B . Steady state SMS mRNA levels relative to GAPDH expression in cultured fibroblasts and osteoblasts. The patient’s cells did not differ significantly from controls. Data were derived by qRT-PCR analysis of 3 independent extractions of total RNA. C . Immunoblot showing steady state SMS protein expression in patient and control osteoblasts. ß-tubulin is shown as a loading control. D . Graph showing steady state SMS protein levels in the patient and control hBMSCs relative to ß-tubulin levels; there was no significant difference. The data are based on 3 independent experiments for each cell line. E - J . Immunofluorescent detection of SMS protein subcellular distribution in unaffected (E-G) and Patient II-1 (H-J) hBMSCs. SMS protein is shown in red and the nucleus is shown in blue. K . Graph quantifying immunoblot detected steady state SMS protein levels in the cytoplasm and nuclei of patient and control hBMSCs. The cytoplasmic expression was normalized to β-tubulin expression and the nuclear expression to p84 expression. L . Polyamine quantification in fibroblasts and osteoblasts. Note that the patient hBMSCs have a more striking imbalance of spermidine and spermine levels than do the patient fibroblasts, * p < 0.05, *** p < 0.005. M . Osteogenic potential of bone marrow stromal cells (hBMSCs) isolated from Patient II-1 sample is markedly lower than that of an unaffected control (cnt). The hBMSCs were seeded in triplicates (6x10 4 /12-well) and either kept untreated (-) or treated (+) with osteogenic differentiation media (see ) for 18 days. After the treatment, cells were fixed and were stained with Alizarin Red S to check for calcium deposition, a marker of osteogenic differentiation.

Article Snippet: We also measured spermine and spermidine levels in cultured human bone marrow stromal cells (hBMSCs) and fibroblasts using the Biochrom 30 polyamine protocol and assessed the osteogenic potential of hBMSCs.

Techniques: Expressing, Cell Culture, Derivative Assay, Quantitative RT-PCR, Western Blot, Control, Isolation, Staining, Marker

Journal: Frontiers in Cellular Neuroscience

Article Title: Bone Marrow Stromal Cells Alleviate Secondary Damage in the Substantia Nigra After Focal Cerebral Infarction in Rats

doi: 10.3389/fncel.2019.00338

Figure Lengend Snippet: Effects of BMSCs treatment on the number of TH, NeuN, and GFAP positive cells in SN, and DA and its metabolites in striatum at 4 weeks after dMCAO. (A) Representative microphotographs of immunohistochemistry for TH (II,VI,X) , NeuN (III,VII,XI) , and GFAP (IV,VIII,XII) in SN. The pictures on the right are magnified from the square area on the left. Scale bar: I, V, IX, 250 μm; II–IV, VI–VIII, X–XII, 50 μm. (B) Quantitative analyses of TH, NeuN, GFAP-positive cells in the SNc at 1 and 4 weeks after dMCAO. Transplantation of BMSCs increased the numbers of TH + [ F (2, 60) = 31.51, p < 0.01] and NeuN + cells [ F (2, 78) = 17.20, p < 0.01], and decreased the number of GFAP + cells [ F (2, 78) = 1848.10, p < 0.01] in the ipsilateral SNc after dMCAO. Each bar represents the mean ± S.D. * p < 0.05 vs. sham-operated group, # p < 0.05 vs. contralateral groups at the same time point and & p < 0.05 vs. ipsilateral vehicle groups ( n = 6 in each group). (C) The DA (I) , DOPAC (II) , and HVA (III) concentrations in striata at 1 and 4 weeks after dMCAO with or without BMSCs transplantation. Transplantation with BMSCs increased the concentration of DA [ F (2, 8) = 6.33, p < 0.05] in the ipsilateral striatum after dMCAO, maintained the concentration of DOPAC [ F (2, 11) = 0.53, p > 0.05] and HVA [ F (2, 11) = 0.67, p > 0.05] in the ipsilateral striatum after dMCAO. Each bar represents the mean ± SD * p < 0.05 vs. sham-operated group and # p < 0.05 vs. contralateral groups at the same time point and & p < 0.05 vs. ipsilateral vehicle groups ( n = 4 in each group). SN, substantia nigra; SNc, substantia nigra compact part; TH, tyrosine hydroxylase; NeuN, neuron-specific nuclear-binding protein; GFAP, Glial fibrillary acidic protein; Sham, sham-operated; BMSCs, bone marrow stromal cells; DA, dopamine; DOPAC, 3,4-dihydroxyphenylacetic acid; HVA, homovanillic acid; con, contralateral; ip, ipsilateral; w, week; dMCAO, distal middle cerebral artery occlusion.

Article Snippet: Over the past two decades, bone marrow stromal cells (BMSCs) based neurorepair has emerged as a promising therapeutic strategy for ischemic stroke.

Techniques: Immunohistochemistry, Transplantation Assay, Concentration Assay, Binding Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Bone Marrow Stromal Cells Alleviate Secondary Damage in the Substantia Nigra After Focal Cerebral Infarction in Rats

doi: 10.3389/fncel.2019.00338

Figure Lengend Snippet: Cortico-striatum-nigral tract retrograde tracing with PRV-152. (A) PRV-152 was injected into the ipsilateral SNr. Regions of interest (ROI) of PRV-152 positive cell counting were shown. Dark shaded area represents ischemic region after dMCAO; lighter shaded area represents ROI. (B) Schematic illustration of the fluorescent signal of PRV-152 in sham-operated and dMCAO groups with or without BMSCs transplantation after PRV-152 was injected into the ipsilateral SNr. (C) Representative photographs of fluorescent double staining of PRV-152 (green) and DAPI (blue) in the ipsilateral cortex (I–IV) , striatum (V–VIII) and SNr (IX-XII) at 4 days after PRV-152 injection in sham-operated group. (D) Representative photographs of fluorescent double staining of PRV-152 (green) and DAPI (blue) in the ipsilateral cortex (I–IV) , striatum (V–VIII) , and SNr (IX–XII) at 4 weeks after PRV-152 injection in vehicle group. (E) Representative photographs of fluorescent double staining of PRV-152 (green) and DAPI (blue) in the ipsilateral cortex (I–IV) , striatum (V–VIII) and SNr (IX–XII) at 4 weeks after PRV-152 injection in BMSCs group. Scale bar, 50 μm. (F) Quantitative analyses of PRV-152 + cell number in the ipsilateral cortex, striatum and SNr after dMCAO. Transplantation with BMSCs increased PRV-152 + cells in the ipsilateral cortex [ F (2, 19) = 9.85, p < 0.01], striatum [ F (2, 41) = 10.67, p < 0.01] and SNr [ F (2, 19) = 6.05, p < 0.01] after dMCAO. Each bar represents the mean ± SD * p < 0.05 vs. sham-operated group and # p < 0.05 vs. vehicle group ( n = 7 in each group). SNr, substantia nigra pars reticulata; Cor, cortex; Str, striatum; PRV, pseudorabies virus; Sham, sham-operated; w, week; dMCAO, distal middle cerebral artery occlusion; BMSCs, bone marrow stromal cells; DAPI, 4’, 6- diamidino-2-phenylindole.

Article Snippet: Over the past two decades, bone marrow stromal cells (BMSCs) based neurorepair has emerged as a promising therapeutic strategy for ischemic stroke.

Techniques: Retrograde Tracing, Injection, Cell Counting, Transplantation Assay, Double Staining, Virus

Journal: International Journal of Nanomedicine

Article Title: Drug Delivery to the Bone Microenvironment Mediated by Exosomes: An Axiom or Enigma

doi: 10.2147/IJN.S307843

Figure Lengend Snippet: Patents Related to the Diagnostic and Therapeutic Application of Exosomes Towards Bone Disorders

Article Snippet: Bone marrow-derived mesenchymal stem cells (BM-MSCs) and Adipose-derived stem cells (ASCs) , WO2019139762A1 , Zen-Bio, Inc. (US) , Exosome compositions and use thereof for joint disorders and diseases , Filed on 2018/Status pending.

Techniques: Diagnostic Assay, Biomarker Discovery, Derivative Assay, CRISPR, Isolation, Injection, Clinical Proteomics

Journal: Korean Journal of Neurotrauma

Article Title: Evaluation of Bone Marrow-derived Stem Cells and Adipose-derived Stem Cells Co-cultured on Human Nucleus Pulposus Cells: A Pilot Study

doi: 10.13004/kjnt.2020.16.e36

Figure Lengend Snippet: SA-β-gal: senescence associated-β-gal, NP: nucleus pulposus, NPC: nucleus pulposus cell, MSC: mesenchymal stem cell, ADMSC: adipose-derived mesenchymal stem cell, BDMSC: bone marrow-derived mesenchymal stem cell.

Article Snippet: BDMSC complete culture medium (ATCC ® PCS-500-030) plus one MSC growth kit (ATCC ® PCS-500-041) and ADMSC complete culture medium (ATCC ® PCS-500-030) plus one MSC growth kit (ATCC ® PCS-500-040) were used to cultivate both MSCs; this medium consists of 485 mL Human MSC Basal Medium, 35 mL Human MSC-Qualified FBS, 0.5 mL penicillin/streptomycin-amphotericin and 6 mL L-alanyl-L-glutamine.

Techniques: Derivative Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Interleukin-37 monomer is the active form for reducing innate immunity

doi: 10.1073/pnas.1819672116

Figure Lengend Snippet: IL-37 elicits a unique (parabolic) dose dependence observed under multiple conditions. (A) Recombinant IL-37 suppression of phorbol 12-myristate 13-acetate (PMA)-induced IL-1β secretion in human peripheral blood mononuclear cells (PBMCs) pretreated with indicated doses of IL-37 2 h before treatment with 25 ng/mL of PMA for 24 h in the presence of 10% FCS. IL-1Ra was also used at 10 μg/mL (B) Recombinant IL-37 (IL-3746–218) suppression of LPS-induced IL-6 secretion in bone marrow cells of mice pretreated with indicated doses of IL-37 2 h before treatment with 100 ng/mL LPS. Assays were performed in quadruplicate in 96 well plates with 500,000 cells/mL. J1 murine macrophage cells were stimulated with LPS, and the indicated doses of IL-3746–218 and supernatants were probed for both (C) IL-1β and (D) IL-6 as described in Fig. 5.

Article Snippet: The murine macrophage cell line J1 was purchased from American Type Culture Collection and cultured according to the distributor’s instructions in DMEM (Corning) supplemented with 10% FBS and 1% penicillin/streptomycin at 37 °C and 5% CO 2 .

Techniques: Recombinant

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Interleukin-37 monomer is the active form for reducing innate immunity

doi: 10.1073/pnas.1819672116

Figure Lengend Snippet: IL-37-mediated antiinflammatory activity is not increased by terminal truncations but only mutations that disrupt dimer formation. (A) J1 murine macrophage cells were pretreated 1 h with the indicated doses of WT IL-3746–218 and engineered terminal mutants of IL-37, stimulated with 1 µg/mL LPS for 4 h and, subsequently, treated with 20 µM of nigericin for 1 h before monitoring IL-1β secretion. (B) WT IL-3746–218 along with both point mutations that specifically disrupt the IL-37 dimer, Y85A, and D73K, was assayed identically to A. Assays were performed in triplicate in a 96 well plate with 750,000 cells/mL Cytokine production was quantified by ELISA. All assays are representative of, at least, five independent experiments. Error bars represent the SEM; *P < 0.05, **P < 0.001, and ***P < 0.0001, statistical significance was assessed using the unpaired Student’s t test.

Article Snippet: The murine macrophage cell line J1 was purchased from American Type Culture Collection and cultured according to the distributor’s instructions in DMEM (Corning) supplemented with 10% FBS and 1% penicillin/streptomycin at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay