Journal: BMC Complementary and Alternative Medicine

Article Title: The effects of Centella asiatica (L.) Urban on neural differentiation of human mesenchymal stem cells in vitro

doi: 10.1186/s12906-019-2581-x

Figure Lengend Snippet: The primer pairs used in the qPCR assay and amplicon sizes

Article Snippet: The primary antibodies used were a mouse monoclonal antibody against S100β (1:1000, Abcam), a rabbit polyclonal antibody against low affinity nerve growth factor receptor (p75 NGFR) (1:1000, Abcam), a mouse monoclonal antibody against myelin binding protein (MBP) (1:1000, Thermo Fisher), a mouse monoclonal antibody against glial fibrillary acidic protein (GFAP) (2 μg/ml, Stem CellTM Technologies) and a mouse monoclonal antibody against myelin oligodendrocyte glycoprotein (MOG) (2 μg/ml, Abcam).

Techniques: Amplification

Journal: BMC Complementary and Alternative Medicine

Article Title: The effects of Centella asiatica (L.) Urban on neural differentiation of human mesenchymal stem cells in vitro

doi: 10.1186/s12906-019-2581-x

Figure Lengend Snippet: Gene expression analysis of S100β, p75 NGFR, MBP and GFAP in differentiated hWJMSCs from different induction groups. Data are presented as relative gene expression normalized to the housekeeping gene, GAPDH. The gene expression level of all the neural gene markers was not significantly different among experimental groups, p < 0.05 ( n = 3). GAPDH; glyceraldehyde 3-phosphate dehydrogenase

Article Snippet: The primary antibodies used were a mouse monoclonal antibody against S100β (1:1000, Abcam), a rabbit polyclonal antibody against low affinity nerve growth factor receptor (p75 NGFR) (1:1000, Abcam), a mouse monoclonal antibody against myelin binding protein (MBP) (1:1000, Thermo Fisher), a mouse monoclonal antibody against glial fibrillary acidic protein (GFAP) (2 μg/ml, Stem CellTM Technologies) and a mouse monoclonal antibody against myelin oligodendrocyte glycoprotein (MOG) (2 μg/ml, Abcam).

Techniques: Gene Expression

Journal: BMC Complementary and Alternative Medicine

Article Title: The effects of Centella asiatica (L.) Urban on neural differentiation of human mesenchymal stem cells in vitro

doi: 10.1186/s12906-019-2581-x

Figure Lengend Snippet: Immunocytochemical analysis of S100β, MBP, GFAP (red) as well as p75 NGFR and MOG (green) in differentiated and undifferentiated hWJMSCs. Cell nuclei were counterstained with DAPI (blue). Undifferentiated hWJMSCs served as a negative control, while Schwann cells acted as a positive control. hWJMSCs differentiated with 1200 μg/ml RECA showed prominent expression of all neural-specific markers compared to differentiated hWJMSCs in other induction groups ( n = 6). DAPI; 4′, 6-diamidino-2-phenylindole. Scale bar 100 μm

Article Snippet: The primary antibodies used were a mouse monoclonal antibody against S100β (1:1000, Abcam), a rabbit polyclonal antibody against low affinity nerve growth factor receptor (p75 NGFR) (1:1000, Abcam), a mouse monoclonal antibody against myelin binding protein (MBP) (1:1000, Thermo Fisher), a mouse monoclonal antibody against glial fibrillary acidic protein (GFAP) (2 μg/ml, Stem CellTM Technologies) and a mouse monoclonal antibody against myelin oligodendrocyte glycoprotein (MOG) (2 μg/ml, Abcam).

Techniques: Negative Control, Positive Control, Expressing

Journal: Frontiers in Immunology

Article Title: IL-1β-Mediated Activation of Adipose-Derived Mesenchymal Stromal Cells Results in PMN Reallocation and Enhanced Phagocytosis: A Possible Mechanism for the Reduction of Osteoarthritis Pathology

doi: 10.3389/fimmu.2019.01075

Figure Lengend Snippet: PMNs reallocate and cluster with ASCs in knees with early CiOA after intra-articular ASC injection. (A,B) Intra-articular injection on day 7 of CiOA resulted in attraction of immune cells within 6 h, both in the control (A) and ASC-injected (B) joints, as was shown in HE-stained total knee joint sections. (C,D) Higher magnifications showed that the immune cells in both saline- (C) and ASC-injected (D) CiOA knee joints had a polymorphonuclear (PMN) phenotype. (E,F) Immunohistochemistry with the specific antibody NIMP-R14 confirmed that a large number of PMNs is attracted to the joints. They were equally spread throughout the synovium and the joint cavity in the control-injected joints (E) , but in the ASC-injected joints a remarkable accumulation of PMNs along the lining was found (F) as indicated by arrows. (G) NIMP-R14 staining showed that 24 h after ASC injection the PMN influx had largely disappeared. (H) ASC injection in naïve joints resulted in NIMP-R14 positive cell influx, albeit lower than in CiOA joints. (I) Quantification of attracted PMNs after intra-articular injection confirmed a significantly increased number of PMNs in CiOA joints compared to naïve joints. (J) At higher magnification we observed a clustering of PMNs around cells we supposed to be ASCs (arrow), which was confirmed with a CD271 antibody (K) which specifically stains ASCs (arrows). Control joints were CiOA knees injected with saline supplemented with 1% BSA. Images shown are representative for the treatment groups ( n = 8 per group). Original magnification × 200 (A,B,E,F,G,H) or × 1,000 (C,D,J,K) . Differences between groups were tested using a one-way ANOVA followed by a Bonferroni Multiple Comparison posttest. F, femur; S, synovium. Bars show mean values ± SD. * P < 0.05.

Article Snippet: The localization of ASCs was visualized with antibodies against CD271 (AP07713PU-N, OriGene).

Techniques: Injection, Control, Staining, Saline, Immunohistochemistry, Comparison

Journal: Cell death & disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells.

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Figure 2 Effects of NGF on BMSCs in monolayer cultures. (a) The MTTassay was used to detect the cytotoxic effects of NGF on BMSCs. (The values are means ± S.D., n = 6, *Po0.05). (b) The effect of NGF on the viability of BMSCs cultured in monolayers for 21 days. Scale bar, 100 μm. (c) Quantification of the DNA content at 7, 14 and 21 days of culture. (d) HE staining was used to determine the morphology of BMCSs cultured in monolayers for 21 days. Scale bar, 100 μm. (e) GAG secretion from the cells after treatment for 7, 14 and 21 days. (f) qRT-PCR was used to determine the expression of the ACAN, SOX9, COL2A1, COL1A1, RUNX2, ENO2, GDNF, BDNF and CNTF genes on days 7, 14 and 21. (g and h) Immunohistochemistry. BMSCs were stained with COL1A1 (g) and COL2A1 (h) antibodies after 21 days in culture. Scale bar, 100 μm. The values are means ± S.D., n = 6; bars with different letters are significantly different from each other at Po0.05

Article Snippet: The proteins were separated by 8% SDS gel electrophoresis and western blotted (N= 10 joints) using antibodies against NGF (Acris OriGene Technologies, Inc., AM05239PU-N, 1:500), COL1A1 (Acris OriGene Technologies, Inc., TA342814, 1:500), COL2A1 (Bioworld Technology, St. Louis Park, MN, USA, BS1071, 1:800), p75NTR (Acris OriGene Technologies, Inc., TA345845, 1:1000), TrkA (Cell Signaling Technology, Danvers, MA, USA, #2505, 1:800), PI3K (Acris OriGene Technologies, Inc., TA330913, 1:1000), AKT (BIORBYT LTD, Cambridge, UK, ORB235003, 1:500), p-Akt (BIORBYT LTD, 05- 802R, 1:800), ERK (Cell Signaling Technology, #4695, 1:800), p-ERK (BIORBYT LTD, T202/Y204, T185/Y187, A303-608A, 1:800), P38 (Acris OriGene Technologies, Inc., TA326350, 1:1000) and p-P38 (BIORBYT LTD, 44-1104G, 1:1000).

Techniques: Cell Culture, Staining, Quantitative RT-PCR, Expressing, Immunohistochemistry

Journal: Cell death & disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells.

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Figure 4 Chondrogenic effects of NGF on BMSCs grown in 3D cultures. (a and b) Immunohistochemical staining of COL1A1 (a) and COL2A1 (b) was performed in the cultured constructs on days 7, 14 and 21. (c) qRT-PCR was performed to determine the expression levels of ACAN, SOX9, COL2A1, COL1A1, RUNX2, ENO2, GDNF, BDNF and CNTF in the cultured constructs on 7 14 and 21 day. (d) Western blots was used to analyze the protein expression levels of Col1A1 and COL2A1. The values are means ± S.D., n = 6; bars with different letters are significantly different from each other at Po0.05, Scale bar = 100 μm

Article Snippet: The proteins were separated by 8% SDS gel electrophoresis and western blotted (N= 10 joints) using antibodies against NGF (Acris OriGene Technologies, Inc., AM05239PU-N, 1:500), COL1A1 (Acris OriGene Technologies, Inc., TA342814, 1:500), COL2A1 (Bioworld Technology, St. Louis Park, MN, USA, BS1071, 1:800), p75NTR (Acris OriGene Technologies, Inc., TA345845, 1:1000), TrkA (Cell Signaling Technology, Danvers, MA, USA, #2505, 1:800), PI3K (Acris OriGene Technologies, Inc., TA330913, 1:1000), AKT (BIORBYT LTD, Cambridge, UK, ORB235003, 1:500), p-Akt (BIORBYT LTD, 05- 802R, 1:800), ERK (Cell Signaling Technology, #4695, 1:800), p-ERK (BIORBYT LTD, T202/Y204, T185/Y187, A303-608A, 1:800), P38 (Acris OriGene Technologies, Inc., TA326350, 1:1000) and p-P38 (BIORBYT LTD, 44-1104G, 1:1000).

Techniques: Immunohistochemical staining, Staining, Cell Culture, Construct, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell death & disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells.

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Figure 6 Chondrogenic effects of NGF on BMSCs in vivo. (a) Safranin O staining were performed on cartilage sections. (b and c) Immunohistochemical staining of COL1A2 (b) and COL1A1 (c) were performed in cartilage sections. (Original low magnification × 40 (scale bar, 500 μm); original high magnification × 200 (scale bar, 100 μm))

Article Snippet: The proteins were separated by 8% SDS gel electrophoresis and western blotted (N= 10 joints) using antibodies against NGF (Acris OriGene Technologies, Inc., AM05239PU-N, 1:500), COL1A1 (Acris OriGene Technologies, Inc., TA342814, 1:500), COL2A1 (Bioworld Technology, St. Louis Park, MN, USA, BS1071, 1:800), p75NTR (Acris OriGene Technologies, Inc., TA345845, 1:1000), TrkA (Cell Signaling Technology, Danvers, MA, USA, #2505, 1:800), PI3K (Acris OriGene Technologies, Inc., TA330913, 1:1000), AKT (BIORBYT LTD, Cambridge, UK, ORB235003, 1:500), p-Akt (BIORBYT LTD, 05- 802R, 1:800), ERK (Cell Signaling Technology, #4695, 1:800), p-ERK (BIORBYT LTD, T202/Y204, T185/Y187, A303-608A, 1:800), P38 (Acris OriGene Technologies, Inc., TA326350, 1:1000) and p-P38 (BIORBYT LTD, 44-1104G, 1:1000).

Techniques: In Vivo, Staining, Immunohistochemical staining