hbmsc complete medium  (Procell Inc)

Journal: PLoS ONE

Article Title: Effects of miR-103 by negatively regulating SATB2 on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells

doi: 10.1371/journal.pone.0232695

Figure Lengend Snippet: Real-time qPCR was performed to detect the mRNA expression levels of RUNX2 (A), BGLAP (B) and SPP1 (C) in HBMScs after 0, 7, 14, and 21 d induction. (D) The expression of miR-103 was markedly reduced in HBMScs during osteogenic differentiation in a time-dependent way. (E and F) Western blot analysis of the protein expression of RUNX, BGLAP and SSP1 in HBMScs during osteogenic differentiation. Columns mean of three independent experiments, and bars SD. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Cells were conventional cultured in OriCell® HBMScs Complete Medium (HUXMA-90011, Cyagen Biosciences) supplemented with 10% of fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and maintained at 37°C in a humidified atmosphere under 5% CO 2 .

Techniques: Expressing, Western Blot

Journal: PLoS ONE

Article Title: Effects of miR-103 by negatively regulating SATB2 on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells

doi: 10.1371/journal.pone.0232695

Figure Lengend Snippet: (A) Elevated expression of miR-103 was determined by real-time qPCR when HBMScs were transfected with agomiR-103. (B) The expression of miR-103 was significantly downregulated in BMMSCs after treated with antagomiR-103. (C) A CCK-8 assay demonstrated that overexpression of miR-103 retarded the growth of HBMScs in vitro . (D) Knockdown of miR-103 promoted HBMScs proliferation ability. Data are shown as the mean ± SD of three independent experiments. * p <0.05, *** p <0.001.

Article Snippet: Cells were conventional cultured in OriCell® HBMScs Complete Medium (HUXMA-90011, Cyagen Biosciences) supplemented with 10% of fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and maintained at 37°C in a humidified atmosphere under 5% CO 2 .

Techniques: Expressing, Transfection, CCK-8 Assay, Over Expression, In Vitro, Knockdown

Journal: PLoS ONE

Article Title: Effects of miR-103 by negatively regulating SATB2 on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells

doi: 10.1371/journal.pone.0232695

Figure Lengend Snippet: (A) Real-time qPCR analysis detected mRNA expression of RUNX2 , BGLAP and SPP1 in HBMScs after treatment with agomiR-103 or agomiR-NC. (B) The RUNX2 , BGLAP and SPP1 mRNA expression were significantly increased in the antagomiR-103 treated HBMScs. (C and D) Western blot analysis of the protein expression of RUNX, BGLAP and SSP1 in HBMScs by miR-103 overexpression and knockdown. (E) The ALP activity of HBMScs was significantly suppressed when miR-103 was overexpressed. (F) The osteogenic differentiation of HBMScs transfected with agomiR-103 and agomiR-NC was observed by Alizarin red S staining. (G) Knockdown of miR-103 exhibited higher ALP activity after osteogenic differentiation of 21 days. (H) Alizarin red S staining investigated the ability of mineralized-bone matrix formation in HBMScs treated with antagomiR-103 or antagomiR-NC. Columns mean of three independent experiments. * p <0.05, *** p <0.001.

Article Snippet: Cells were conventional cultured in OriCell® HBMScs Complete Medium (HUXMA-90011, Cyagen Biosciences) supplemented with 10% of fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and maintained at 37°C in a humidified atmosphere under 5% CO 2 .

Techniques: Expressing, Western Blot, Over Expression, Knockdown, Activity Assay, Transfection, Staining

Journal: PLoS ONE

Article Title: Effects of miR-103 by negatively regulating SATB2 on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells

doi: 10.1371/journal.pone.0232695

Figure Lengend Snippet: (A) miR-103 target sites in the 3’-UTR of SATB2 mRNA. (B) Real-time qPCR and ELISA were conducted to examine the effects of miR-103 overexpression on the mRNA and protein expression of SATB2. GAPDH was also detected as a loading control. (C) Knockdown of miR-103 markedly upregulated SATB2 mRNA and protein expression in HBMScs. (D) Relative luciferase activity of the WT or MUT SATB2 reporter plasmids along with agomiR-103 or agomiR-NC in HBMScs was shown. Renilla luciferase activity was normalized to Firefly luciferase activity and plotted as relative luciferase activity. (E) Luciferase activities were measured in HBMScs following cotransfection with the WT or MUT SATB2 reporter plasmids and antagomiR-103 or antagomiR-NC. Data are shown as the mean ± SD of three independent experiments. ** p <0.01, *** p <0.001.

Article Snippet: Cells were conventional cultured in OriCell® HBMScs Complete Medium (HUXMA-90011, Cyagen Biosciences) supplemented with 10% of fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and maintained at 37°C in a humidified atmosphere under 5% CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Over Expression, Expressing, Control, Knockdown, Luciferase, Activity Assay, Cotransfection

Journal: PLoS ONE

Article Title: Effects of miR-103 by negatively regulating SATB2 on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells

doi: 10.1371/journal.pone.0232695

Figure Lengend Snippet: (A) Expression levels of SATB2 mRNA and protein were examined after transfection of SATB2 siRNA and siRNA NC. (B) Transfection of SATB2 siRNA partly reversed the upregulation of SATB2 expression by antagomiR-103. (C) HBMScs co-transfected with antagomiR-103 and SATB2 siRNA exhibited a lower cell proliferation ability than cells transfected with antagomiR-103 and siRNA NC. (D) SATB2 knockdown partly abolished antagomiR-103 induced ALP activity of HBMScs. (E) SATB2 knockdown partly abolished antagomiR-103 induced mineralized matrix formation of HBMScs. (F and G) HBMScs treatment with antagomiR-103 and SATB2 siRNA reduced the protein expression levels of RUNX2, BGLAP, and SPP1 compared cells treated with antagomiR-103 and siRNA NC. Columns mean of three independent experiments. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Cells were conventional cultured in OriCell® HBMScs Complete Medium (HUXMA-90011, Cyagen Biosciences) supplemented with 10% of fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and maintained at 37°C in a humidified atmosphere under 5% CO 2 .

Techniques: Expressing, Transfection, Knockdown, Activity Assay