msc basal medium + supplements  (AllCells LLC)

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Extract (MSCsE)-Based Therapy Alleviates Xerostomia and Keratoconjunctivitis Sicca in Sjogren’s Syndrome-Like Disease

doi: 10.3390/ijms20194750

Figure Lengend Snippet: Salivary and lacrimal glands function represented as SFR (Salivary Flow Rate) and TFR (Tear Flow Rate), respectively. SFR and TFR were assessed pre-treatment at week 0 (8-week-old) then 4, 8, 12, and 16 weeks post-treatment. ( A ) SFR was determined by volume of saliva/min/gm body weight (multiplied by 10 for simplicity). Control group showed a continuous decrease of SFR (lost almost 47% of SFR at week 16 in comparison to the highest reached level, week 4) whereas MSCs-/MSCsE-treated groups maintained a significantly higher SFR (maintained almost 75–100% of SFR at week 4) than the control, their results were comparable to each other, and to the ICR group, ( n = 5–12). ( B ) TFR was determined by length of wetted phenol red thread in mm/min. Control group showed a continuous decrease of TFR; whereas, MSCs-/MSCsE-treated groups maintained significantly higher TFRs that are comparable to each other and to the wild type ICR group. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001, n = 3–6. All data were presented as mean ± S.D. Control: saline-treated; MSCs: Mesenchymal stem cells; MSCsE: Mesenchymal stem cells extract.

Article Snippet: The BM pellet was reconstituted and mixed thoroughly with the MSCs media (MesenCultTM MSC Basal Medium+ MesenCultTM Mesenchymal Stem Cell Stimulatory Supplements (STEMCELL)).

Techniques: Control, Comparison, Saline

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Extract (MSCsE)-Based Therapy Alleviates Xerostomia and Keratoconjunctivitis Sicca in Sjogren’s Syndrome-Like Disease

doi: 10.3390/ijms20194750

Figure Lengend Snippet: Special cell subpopulations in submandibular (SMG) and lacrimal glands (LG) were evaluated by immunofluorescence staining at 16 weeks post-treatment. ( A , C ) SMG/LG immunofluorescence staining, respectively, positive for AQP5 (marker for water channel protein expressed by acinar cells in SMG and acinar/ductal cells in LG), α-SMA (marker for myoepithelial cells), AQP4 (marker for acinar and ductal cells), CK5 (marker for ductal/progenitor cells), and c-Kit (marker for stem/progenitor cells) were tested in frozen sections, scale bar = 148 μm. (B , D ) Quantification of protein immunofluorescence expression levels in submandibular/lacrimal glands, respectively, from 4–6 random fields/glands by Image J software. MSCs-/MSCsE-treated groups showed higher intensities for all the tested markers when compared with the control group. All images were randomly taken at 200× magnification. * p ≤ 0.05; ** p ≤ 0.01, n = 3–6. All data were presented as mean ± S.D. Control: saline-treated; MSCs: Mesenchymal stem cells; MSCsE: Mesenchymal stem cells extract.

Article Snippet: The BM pellet was reconstituted and mixed thoroughly with the MSCs media (MesenCultTM MSC Basal Medium+ MesenCultTM Mesenchymal Stem Cell Stimulatory Supplements (STEMCELL)).

Techniques: Immunofluorescence, Staining, Marker, Expressing, Software, Control, Saline

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Extract (MSCsE)-Based Therapy Alleviates Xerostomia and Keratoconjunctivitis Sicca in Sjogren’s Syndrome-Like Disease

doi: 10.3390/ijms20194750

Figure Lengend Snippet: Proliferation rates, serum EGF levels, and gene expression levels of key genes at 16 weeks post-treatment. ( A ) Immunofluorescence staining of submandibular (SMG) (upper panel) and lacrimal (LG) (lower panel) glands for proliferation protein Ki-67 (red), nuclei were stained with DAPI (blue), ( B ) Proliferation rate (Ki-67-positive cells %) for submandibular and lacrimal glands was calculated using random 200× magnified images (acquired by Volocity software) using Image J software. Two examiners independently analyzed images in a blind manner. MSCs/MSCsE treatments promoted tissue proliferation in the glands significantly higher than the control group and their rates were comparable to the ICR group. ( C ) Serum levels of EGF measured by ELISA. Both treatments elevated EGF levels in comparison to the control group, but only MSCsE treatment induced a significantly higher level. ( D ) Relative expression of key genes for tissue function, repair, regeneration, and apoptosis were analyzed by quantitative RT-PCR in lacrimal (LG) and submandibular (SG) glands. Gene expression levels in the lacrimal gland were significantly higher in MSCs-/MSCsE-treated groups than that of the control for AQP5, EGF, LYZ1, MMP2, and BMP7. Gene expression levels in the submandibular gland were significantly higher in MSCs-/MSCsE-treated groups than that of the control for AQP5, EGF, FGF2, and BMP7; and significantly lower for MMP2 and CASP3. Y-axis shows the relative expression of the gene compared to GAPDH, three experimental replicates were conducted for each sample. Scale bar = 74 μm, * p < 0.05, ** p < 0.01; *** p < 0.001, n = 3–6. All data were presented as mean ± S.D. Control: saline-treated; MSCs: Mesenchymal stem cells; MSCsE: Mesenchymal stem cells extract.

Article Snippet: The BM pellet was reconstituted and mixed thoroughly with the MSCs media (MesenCultTM MSC Basal Medium+ MesenCultTM Mesenchymal Stem Cell Stimulatory Supplements (STEMCELL)).

Techniques: Gene Expression, Immunofluorescence, Staining, Software, Control, Enzyme-linked Immunosorbent Assay, Comparison, Expressing, Quantitative RT-PCR, Saline

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Extract (MSCsE)-Based Therapy Alleviates Xerostomia and Keratoconjunctivitis Sicca in Sjogren’s Syndrome-Like Disease

doi: 10.3390/ijms20194750

Figure Lengend Snippet: Thickness of the central cornea (total) and the corneal epithelium at 16 weeks post-treatment. ( A ) H&E stained images of the cornea, arrowheads represent the epithelial thickness. ( B ) Analysis of the total cornea thickness. ( C ) Analysis of corneal epithelium thickness. Images of H&E stained sections of the cornea were obtained using Volocity software then Image J was used to assess the thickness. The MSCs-/MSCsE-treated groups showed a significantly higher corneal epithelial thickness. Scale bar = 74 μm, * p ≤ 0.05, n = 3–6. All data were presented as mean ± S.D. Control: saline-treated; MSCs: Mesenchymal stem cells; MSCsE: Mesenchymal stem cells extract.

Article Snippet: The BM pellet was reconstituted and mixed thoroughly with the MSCs media (MesenCultTM MSC Basal Medium+ MesenCultTM Mesenchymal Stem Cell Stimulatory Supplements (STEMCELL)).

Techniques: Staining, Software, Control, Saline

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Extract (MSCsE)-Based Therapy Alleviates Xerostomia and Keratoconjunctivitis Sicca in Sjogren’s Syndrome-Like Disease

doi: 10.3390/ijms20194750

Figure Lengend Snippet: Focus score, focus area and lymphocytes composition analysis in the submandibular (SMG) and lacrimal (LG) glands at 16 weeks post-treatment. ( A ) H&E stained images of lymphocytic infiltrates in the submandibular glands (upper panel) and lacrimal glands (lower panel). ( B ) Focus score analysis (number of lymphocytic infiltrates/4 mm 2 ) using serial H&E stained sections, cut at different levels, under the light microscope. The analysis revealed a lower score for the treated groups but was not statistically significant. ( C ) Focus area (in µm 2 ) was calculated by Image J software using H&E images (400×/200×) acquired by Volocity software. Treated groups showed significantly smaller focus areas. Immunohistochemical staining of lymphocytic infiltrate for B220 (a pan B cell marker in mice), BAFF (B cells activating factor), and FoxP3 (forkhead box P3, T reg marker) in submandibular glands ( D ) and lacrimal glands ( E ). ( F , G ) Quantification of protein expression for BAFF and B220, respectively. The positive signals were measured using Image J software then divided by the size of the lymphocytic infiltrate (focus area). The results were represented as % of signal intensity. ( H ) Quantification of FoxP3 + T reg cells. Positive cells were counted in each lymphocytic infiltrate then divided by the focus area (cell/ µm 2 ) using Image J software. MSCs/MSCsE groups exhibited a significantly higher FoxP3 + and lower B220 + and BAFF + cells in the lymphocytic infiltrates when compared to the control group. All images were taken at 200× magnification. Scale bar = 148 μm, * p ≤ 0.05; ** p ≤ 0.01, n = 3–6. All data were presented as mean ± S.D. Control: saline-treated; MSCs: Mesenchymal stem cells; MSCsE: Mesenchymal stem cells extract.

Article Snippet: The BM pellet was reconstituted and mixed thoroughly with the MSCs media (MesenCultTM MSC Basal Medium+ MesenCultTM Mesenchymal Stem Cell Stimulatory Supplements (STEMCELL)).

Techniques: Staining, Light Microscopy, Software, Immunohistochemical staining, Marker, Expressing, Control, Saline

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Extract (MSCsE)-Based Therapy Alleviates Xerostomia and Keratoconjunctivitis Sicca in Sjogren’s Syndrome-Like Disease

doi: 10.3390/ijms20194750

Figure Lengend Snippet: Serum levels of anti-SSA/Ro autoantibodies and IL-10, and gene expression levels of anti-/pro-inflammatory cytokines/factors at 16 weeks post-treatment. ( A ) Serum levels of anti-SSA/Ro autoantibodies (assessed by ELISA) for MSCs-/MSCsE-treated groups exhibited significantly lower levels in comparison to the control group. ( B ) Serum levels of IL-10 (assessed by ELISA) for MSCs-/MSCsE-treated groups were significantly higher than the control group. ( C ) Gene expression levels for anti-/pro-inflammatory cytokines/factors were measured using quantitative RT-PCR in the submandibular and lacrimal glands. MSCs/MSCsE treatments upregulated IL-10 and down-regulated TNF-α gene expressions in both glands, and down-regulated TGF-β, IL-1β in lacrimal glands. Y-axis shows the relative expression of the gene compared to GAPDH, three experimental replicates were conducted for each sample. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p ≤ 0.0001, n = 4–6. All data were presented as mean ± S.D. Control: saline-treated; MSCs: Mesenchymal stem cells; MSCsE: Mesenchymal stem cells extract.

Article Snippet: The BM pellet was reconstituted and mixed thoroughly with the MSCs media (MesenCultTM MSC Basal Medium+ MesenCultTM Mesenchymal Stem Cell Stimulatory Supplements (STEMCELL)).

Techniques: Gene Expression, Enzyme-linked Immunosorbent Assay, Comparison, Control, Quantitative RT-PCR, Expressing, Saline