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: Journal of Cellular and Molecular Medicine

Article Title: Human Decidua Basa lis mesenchymal stem/stromal cells reverse the damaging effects of high level of glucose on endothelial cells in vitro

doi: 10.1111/jcmm.15248

Figure Lengend Snippet: Measurement of HUVEC migration under various treatment conditions, using xCELLigence Real‐Time Cell Analyser. Different HUVEC treatment groups used in the migration experiments included: HUVEC cultured alone (A), HUVEC cultured with 100 mmol/L glucose (B), HUVEC cultured with 100 mmol/L glucose and 25% DBMSC conditioned medium (CMDBMSC) (C) in the upper chamber of the CIM migration plate, whereas HUVEC medium with 30% FBS was added to the lower chamber. HUVEC cultured in the upper chamber, whereas 100 mmol/L glucose was added to the lower chamber (D), HUVEC cultured in the upper chamber, whereas 100 mmol/L glucose and 25% CMDBMSC were added to the lower chamber (E). Pre‐Glu (HUVEC pre‐treated with 100 mmol/L glucose for 72 hours, harvested and seeded in the upper chamber) (F), Pre‐CM (HUVEC pre‐treated with 100 mmol/L glucose and 25% CMDBMSC for 72 hours, harvested and seeded in the upper chamber) (G), and Pre‐IC (HUVEC pre‐treated with 100 mmol/L glucose and DBMSCs at a ratio of 1HUVEC:1DBMSC in a cellular communication culture system (ICDBMSC) for 72 hours, harvested and seeded in the upper chamber) (H). Migration of HUVEC cultured with 100 mmol/L glucose (100 (in) as compared to glucose‐untreated HUVEC (I). HUVEC migration in presence of 100 mmol/L glucose and CMDBMSC [100 + CM (in)] (I). HUVEC migration in response to 100 mmol/L glucose alone [100 (out)], added to the lower chamber of the migration plate, as compared to glucose‐untreated HUVEC after 24 hours (J). Migration of HUVEC pre‐treated with glucose (Pre‐Glu), or with 100 mmol/L glucose and CMDBMSC (Pre‐CM) compared to glucose‐untreated HUVEC (K). Each experiment was performed in triplicate using HUVEC (passage 3‐5) and DBMSCs (passage 3) from five independent umbilical cord tissues, and placentae, respectively. Bars represent standard errors

Article Snippet: MSCs were isolated from the decidua basalis (DBMSCs) of human term placenta, whereas HUVEC (human umbilical vein endothelial cells) were isolated from umbilical cord veins as previously described by us., DBMSCs were cultured in complete DBMSC culture medium [DMEM‐F12 medium containing 10% foetal bovine serum (MSC‐FBS, catalogue number 12‐662‐011, Life Technologies), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], whereas HUVEC were cultured in complete endothelial cell growth medium (Catalogue number PCS‐100‐041 TM , ATCC).

Techniques: Migration, Cell Culture

Journal: Journal of Cellular and Molecular Medicine

Article Title: Human Decidua Basa lis mesenchymal stem/stromal cells reverse the damaging effects of high level of glucose on endothelial cells in vitro

doi: 10.1111/jcmm.15248

Figure Lengend Snippet: The proliferation of HUVECs in response to 100 mmol/L glucose and different treatments of DBMSCs. MTS assay showed that proliferation of HUVEC in response to 100 mmol/L glucose significantly reduced as compared to glucose‐untreated HUVEC controls after 72 hours, whereas treatment with 25% CMDBMSC (100 + CM) and 100 mmol/L glucose for 72 hours significantly increased HUVEC proliferation as compared to glucose‐untreated and treated HUVECs. HUVEC proliferation in response to 100 mmol/L glucose and DBMSCs (100 + DBMSC) significantly reduced as compared to glucose‐untreated HUVEC and did not significantly change as compared to glucose‐treated HUVEC after 72 hours. Each experiment was performed in triplicate using DBMSCs (passage 3) and HUVEC (passage 3‐5) from five independent placentae and umbilical cord tissues. * P < .05. Bars represent standard errors

Article Snippet: MSCs were isolated from the decidua basalis (DBMSCs) of human term placenta, whereas HUVEC (human umbilical vein endothelial cells) were isolated from umbilical cord veins as previously described by us., DBMSCs were cultured in complete DBMSC culture medium [DMEM‐F12 medium containing 10% foetal bovine serum (MSC‐FBS, catalogue number 12‐662‐011, Life Technologies), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], whereas HUVEC were cultured in complete endothelial cell growth medium (Catalogue number PCS‐100‐041 TM , ATCC).

Techniques: MTS Assay

Journal: Scientific Reports

Article Title: Bile acids induce hepatic differentiation of mesenchymal stem cells

doi: 10.1038/srep13320

Figure Lengend Snippet: ( a,b ) Freshly isolated rat HSCs were treated with 2 μM TLCA, TCDCA, TCA, GCDCA, CA or TUDCA for 21 days in serum-free media (n = 3). The HSCs were analysed in weekly intervals by qPCR of ( a ) albumin and ( b ) Hnf4α mRNA. Hprt1 expression was used for the normalization of the values and the highest mRNA amount of the hepatic markers measured by qPCR was set to 100%. The gradual increase of the hepatocyte markers albumin and Hnf4α in HSC cultures indicated that stellate cells differentiated into liver parenchymal cells in response to bile acid treatment. In contrast to this, HSC of the control, which were treated with serum-free medium without bile acids, remained negative for hepatocyte markers. ( c,d ) Freshly isolated HSC and ( e,f ) cultured bmMSC from rats were treated with 2 μM TUDCA for 21 days (n = 3). ( c,e ) The hepatocyte marker albumin was quantified at the protein level by ELISA in culture supernatants. ( d,f ) The release of bile acids into the culture medium was measured by UHPLC-MS/MS in weekly intervals. Hepatic differentiation as indicated by induced albumin expression and bile acid synthesis was congruently initiated in rat HSC and bmMSC of rats by TUDCA treatment, whereas MSC populations without TUDCA (control) remained negative for these hepatocyte markers. TUDCA levels close to and above 2 μmol/L were also measured by UHPLC-MS/MS. Since TUDCA was added to the medium, it was not included in the graphs. ( d,f ) Small amounts of bile acids were already found in the culture medium by UHPLC-MS/MS before incubation of cells, which may account for the low concentrations of bile acids measured at day 1 and under control conditions.

Article Snippet: Bone marrow cells that remained unselected by this method were seeded on plastic dishes coated with collagen type 1 (#356401, BD Biosciences, Heidelberg, Germany), and cultured in MSC expansion medium (#CCM004; StemXVivo Mesenchymal Stem Cell Expansion Medium; R&D Systems, Minneapolis, MN, USA).

Techniques: Isolation, Expressing, Control, Cell Culture, Marker, Enzyme-linked Immunosorbent Assay, Tandem Mass Spectroscopy, Incubation

Journal: OncoTargets and therapy

Article Title: TGFβ1 is essential for MSCs-CAFs differentiation and promotes HCT116 cells migration and invasion via JAK/STAT3 signaling

doi: 10.2147/OTT.S178618

Figure Lengend Snippet: The effects of conditioned medium of colorectal cancer cells on the differentiation of MSCs to CAFs. ( A ) After HCT116 and HT29cells were cultured with serum-free medium for 48 hrs, the media was saved to perform TGFβ1, TNFα, HGF and PDGF levels via Enzyme-linked immunosorbent assay (ELISA). * P <0.05 and ** P <0.01 vs Con-CM. ( B ) The condition of MSCs cultured under serum-free medium (Con-CM) or HCT116 or HT29 conditioned medium (HCT116-CM or HT29-CM) for 48 hrs. Scale bar, 500 µm. ( C ) CCK-8 assay to detect the proliferation of MSCs for indicated times. * P <0.05 and ** P <0.01 vs Con-CM. ( D ) Sphere formation assay of MSCs incubated with Con-CM and HCT116-CM or HT29-CM. Scale bar, 500 µm. ( E ) For flow cytometry, the CAFs-related biomarkers such as α-SMA, vimentin, FSP1 were detected. The experiments were repeated at least three times. Abbreviations: Con, control; MSCs, mesenchymal stem cells; CAFs, cancer-associated fibroblasts.

Article Snippet: The human colorectal carcinoma cell lines (HCT116, HT29) and bone-marrow-derived MSCs were obtained from ATCC.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Tube Formation Assay, Incubation, Flow Cytometry, Control

Journal: OncoTargets and therapy

Article Title: TGFβ1 is essential for MSCs-CAFs differentiation and promotes HCT116 cells migration and invasion via JAK/STAT3 signaling

doi: 10.2147/OTT.S178618

Figure Lengend Snippet: TGFβ1-blocking peptide P17 suppresses the proliferation and differentiation of MSCs to CAFs. ( A ) P17 was pre-treated colorectal cancer cells and then the condition of MSCs incubated with various conditioned medium was detected. Scale bar, 500 µm. ( B ) Proliferation of MSCs was detected by CCK8 assay. ( C ) The ability of MSCs to form spheres in Con-CM group and HCT116-CM+P17 or HT29+P17 group was inhibited compared with HCT116-CM or HT29-CM group. ** P <0.01 vs Con-CM, ** P <0.01 vs HCT116-CM and ** P <0.01 vs HT29-CM group. ( D ) The expression of CAFs-related proteins such as α-SMA, vimentin, FSP was measured by flow cytometry. All experiments were at least repeated three times. Abbreviations: Con, control; MSCs, mesenchymal stem cells; CAFs, cancer-associated fibroblasts; CM, conditioned medium.

Article Snippet: The human colorectal carcinoma cell lines (HCT116, HT29) and bone-marrow-derived MSCs were obtained from ATCC.

Techniques: Blocking Assay, Incubation, CCK-8 Assay, Expressing, Flow Cytometry, Control

Journal: OncoTargets and therapy

Article Title: TGFβ1 is essential for MSCs-CAFs differentiation and promotes HCT116 cells migration and invasion via JAK/STAT3 signaling

doi: 10.2147/OTT.S178618

Figure Lengend Snippet: TGFβ1 induces HCT116 and HT29 cells migration and invasion via activating JAK/STAT3 signaling. ( A – D ) The migration and invasion of HCT116 cells was determined by wound-healing and transwell assays. * P <0.05 and ** P <0.01 vs Con-CM, * P <0.05 and ** P <0.01 vs Con-CM+TGFβ1, ** P <0.01 vs HCT116-CM or HT29-CM. ( E ) p-STAT3 protein levels in HCT116 cells subcellular fractions with TGFβ1, P17 or AG490. GAPDH and Lamin A were used to act as cytoplasmic and nuclear markers, respectively. ( F ) The molecular pathway of TGFβ1 inducing MSCs-CAFs' differentiation and CRC metastasis. Abbreviations: Con, control; CM, conditioned medium; MSCs; mesenchymal stem cells; CAFs, cancer-associated fibroblasts; CRC, colorectal cancer.

Article Snippet: The human colorectal carcinoma cell lines (HCT116, HT29) and bone-marrow-derived MSCs were obtained from ATCC.

Techniques: Migration, Control