Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: DBMSC proliferation groups. A Group 1 consisted of DBMSC cultured alone in a complete DBMSC culture medium. B Group 2 consisted of DBMSC cultured with different concentrations (25–400 mM) of glucose in a complete DBMSC culture medium. C Group 3 consisted of DBMSC pretreated with 200 mM glucose for 72 h [200 (pre)], harvested and then re-cultured alone in a complete DBMSC culture medium. D DBMSCs were seeded in a 16-well plate (E-Plate 16). The culture plates were then placed in the xCELLigence system at 37 °C in a cell culture incubator, and DBMSC cell index was then monitored. DBMSC proliferation in response to different glucose concentrations by the xCELLigence system. As compared to untreated DBMSCs, DBMSC proliferation was unchanged at 25 mM glucose ( p > 0.05) but significantly increased at 50 and 200 mM glucose and then significantly reduced at 400 mM glucose, after 24 h in culture. E At 48 h in culture, and as compared to untreated DBMSCs, DBMSC proliferation was unchanged at 50 mM glucose ( p > 0.05) but significantly increased at 200 mM glucose and then significantly reduced at 25 and 400 mM glucose. F At 72 h in culture, and as compared to untreated DBMSCs, DBMSC proliferation significantly increased at 200 mM glucose but was significantly reduced at 25, 50 and 400 mM glucose. G - I The reversibility of DBMSC proliferation in response to glucose. DBMSCs were initially cultured with 200 mM glucose for 72 h and their proliferation was then determined using the xCELLigence system. At 24–72 h in culture, and as compared to untreated DBMSCs and DBMSC-treated with 200 mM glucose [200 (I)], the proliferation of DBMSC pretreated with 200 mM glucose [200 (pre)] significantly reduced

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Cell Culture

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: DBMSC migration groups. A Group 1 consisted of DBMSCs cultured alone in the upper chamber. B Group 2 consisted of DBMSCs cultured alone in the upper chamber while 200 mM glucose was added to the lower chamber. C Group 3 consisted of DBMSCs pre-treated with 200 mM glucose for 72 h (200(pre)) harvetsed and then re-cultured alone in the upper chamber while 200 mM glucose was added to the lower chamber. D DBMSCs were seeded in DBMSC serum free medium in the upper chamber of the CIM migration plate while DBMSC culture medium containing 30% FBS was added to the lower chambers. At 24 h, DBMSC [DB (T 200)] migration in response to 200 mM glucose significantly increased as compared to untreated DBMSCs (DB). The migration of DBMSCs pretreated with 200 mM glucose for 72 h (Pre-DB) in response to 200 mM glucose [Pre-DB (To 200)] significantly increased as compared to untreated DBMSCs (DB), but was unchanged as compared to DBMSC migrating in response to 200 mM glucose [DB (To 200)], p > 0.05. E The effect of glucose on DBMSC invasion through endothelial cells by the xCELLigence system. At 10 h, the pretreatment with 200 mM glucose for 72 h [200 (Pre)] significantly increased DBMSC invasion as compared to untreated DBMSCs and DBMSC cultured with 200 mM glucose while the addition of 200 mM glucose [200 (in)] during the invasion experiment had no significant effect on DBMSC invasion as compared to untreated DBMSCs. Each experiment was performed in triplicate and repeated with five independent DBMSC (passage 3) preparations. * p < 0.05. Bars represent standard errors

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Migration, Cell Culture

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: BMSCs were cultured with 200 mM glucose [200 (I)] and their adhesion was then determined using the xCELLigence system. As compared to untreated DBMSCs, the treatment with 200 mM glucose had no significant effect on DBMSC adhesion at 2 h in culture ( p > 0.05) while the adhesion of DBMSC pretreated with 200 mM glucose for 72 h was significantly increased at 2 h as compared to untreated DBMSCs, and DBMSC cultured with 200 mM glucose [200 (I)]. Each experiment was performed in triplicate and repeated with five independent DBMSC (passage 3) preparations. * p < 0.05. Bars represent standard errors

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Cell Culture

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Flow cytometric analysis of DBMSC expression of immune markers. A - C The treatment with 200 mM glucose significantly increased the DBMSCs [200 (pre)] expression of ICAM-1, had no significant effect on IL-12 expression, p > 0.05, and significantly increased the expression of B7H4, as compared untreated DBMSCs. Each experiment was performed in triplicate and repeated with five independent DBMSC (passage 3) preparations. * p < 0.05. Bars represent standard errors

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes with survival, anti-apoptotic, proliferation, and migration properties. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing, Migration

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes with pro-oxidant and antioxidant properties. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose increased DBMSC expression of genes with antioxidant, anti-inflammatory, anti-chemoattractant, and antimicrobial properties. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing, Clinical Proteomics

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing, Membrane

Journal: Nature Communications

Article Title: p16 INK4A -deficiency predicts response to combined HER2 and CDK4/6 inhibition in HER2+ breast cancer brain metastases

doi: 10.1038/s41467-022-29081-2

Figure Lengend Snippet: a Genomic analysis of genes associated with cell cycle pathway in BCBM PDXs. Major genetic alterations across 30 BCBM PDX samples. b CDKN2A mRNA expression levels across BCBM PDXs. Each dot represents one PDX model: HER2+/ER− ( n = 10), HER2+/ER+ ( n = 10), ER+/HER2− ( n = 8), triple negative (TN) ( n = 8), mean ± SD, one-way ANOVA followed by Tukey’s multiple comparisons test. c Heatmap of CDKN2A and RB1 mRNA levels across BCBM PDXs. d Bar plot of p16 INK4A positive cells by immunohistochemistry (IHC) per HER2 + PDX model with CDNK2A copy number variation (CNV) shown below. p16 INK4A positive cells shown as mean ± SD of three images per section per sample. e p16 INK4A IHC of the indicated BCBM PDXs (Scale bar, 50 μm). HCC70 xenograft tumor sections were used as a positive control. f Comparison of p16 INK4A positive cells in HER2 + BCBM PDX and matched patient samples ( n = 11, mean ± SD). Mean PDX p16 INK4A positive cells 9.7 ± 3.4%, mean patient p16 INK4A positive cells 33.6 ± 8.9%, p = 0.06. ns: not significant. g Proportion of patients in our BCBM cohort either negative or positive for p16 INK4A expression based on 30% cutoff. Source data are provided as a Source Data file.

Article Snippet: Overexpression of p16 INK4A in cells were generated by infecting the cells with lentivirus encoding from pLX401-INK4A (Addgene #121919, doxycycline-inducible p16 INK4A expression).

Techniques: Expressing, Immunohistochemistry, Positive Control, Comparison

Journal: Nature Communications

Article Title: p16 INK4A -deficiency predicts response to combined HER2 and CDK4/6 inhibition in HER2+ breast cancer brain metastases

doi: 10.1038/s41467-022-29081-2

Figure Lengend Snippet: a Bioluminescence imaging (BLI) analysis with ROI of mice bearing DFBM-727 (p16 INK4A positive) tumors at indicated weeks after treatment with control, tucatinib (PO 75 mg/kg, BID), abemaciclib (PO 75 mg/kg, QD), or Tuca + Abem. Left panel: representative bioluminescence images. Middle panel: quantification of the regions of interest (ROI) in each group mice at indicated imaging time points, Vehicle control ( n = 5), Tucatinib ( n = 4), Abemaciclib (wk0-4, n = 4; wk6, n = 2) or Tuca + Abem ( n = 5). Mean ± SD. Right panel: Kaplan–Meier survivals of mice bearing DFBM-727. Dotted lines indicate treatment starting time. b BLI and survival analysis for DFBM-Ni8 (p16 INK4A IHC negative, CDKN2A CNV = 1) treated as in a . BLI right panel: Vehicle control (wk0,1 n = 5; wk2,3, n = 4; wk4, n = 3), Tucatinib ( n = 5), Abemaciclib (wk0-3, n = 4; wk4, n = 3) or Tuca + Abem (wk0-2, n = 5; wk3,4, n = 4). Mean ± SEM. c BLI and survival analysis for DFBM-Ni17 (p16 INK4A IHC negative, CDKN2A CNV = 2) treated as in a . BLI right panel: Vehicle control ( n = 3), Tucatinib ( n = 3), Abemaciclib (wk0, n = 3; wk1,2, n = 2), or Tuca + Abem ( n = 3). Mean ± SEM. d Multiplex immunofluorescence shows increase in p16 INK4A (control, n = 8 sections; Dox, n = 7 sections) and reduction in p-RB (control, n = 7 sections; Dox, n = 8 sections) after 3 days of doxycycline administration to DFBM-Ni17/tetp16 INK4A . Mean ± SD, Unpaired t -test. e BLI analysis with ROI of mice bearing DFBM-Ni17/tetp16 INK4A tumors at the indicated time with the treatment as indicated. Tucatinib (PO 75 mg/kg, BID), abemaciclib (PO 75 mg/kg, QD), doxycycline-diet (2500 ppm, ScottPharma). Control ( n = 4), Dox ( n = 4), Dox + Abem ( n = 4), Dox + Tuc (wk0-3, n = 5; wk5, n = 4), Dox + Tuc + Abem (wk0-1, n = 5; wk2-5, n = 4). Mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Overexpression of p16 INK4A in cells were generated by infecting the cells with lentivirus encoding from pLX401-INK4A (Addgene #121919, doxycycline-inducible p16 INK4A expression).

Techniques: Imaging, Control, Multiplex Assay, Immunofluorescence

Journal: Nature Communications

Article Title: p16 INK4A -deficiency predicts response to combined HER2 and CDK4/6 inhibition in HER2+ breast cancer brain metastases

doi: 10.1038/s41467-022-29081-2

Figure Lengend Snippet: Schematic diagram of rational combination of tucatinib and abemaciclib for HER2+, p16 INK4A -deficient BCBMs.

Article Snippet: Overexpression of p16 INK4A in cells were generated by infecting the cells with lentivirus encoding from pLX401-INK4A (Addgene #121919, doxycycline-inducible p16 INK4A expression).

Techniques: