xenograft pancreatic cancer stem cells cscs  (Celprogen Inc)

Journal: Journal of materials chemistry. B

Article Title: Design and evaluation of nanoscale materials with programmed responsivity towards epigenetic enzymes.

doi: 10.1039/d4tb00514g

Figure Lengend Snippet: Fig. 6 Interaction of the block copolymers and HDAC8-responsive nanoparticles with different types of PDAC cells and non-cancerous HPNE cells in vitro. (A) PEG-block-poly(acetylated L-lysine) block copolymers studied in this work do not trigger cytotoxicity in different cell lines. (B) Chemical structure of the STAT3 inhibitor, Napabucasin (NAPA), which has been used as the model hydrophobic drug to demonstrate the encapsulation and HDAC-mediated release activity of the nanoparticles in the context of drug delivery. (C) HDAC 8 (1 mM) triggers the release of NAPA, and over 90% of the encapsulated drug is released from these nanoparticles after incubation with the enzyme for 3 h. The standard deviation of mean is taken for N = 3 replicates. Without the enzyme, the drug release rate and extent were significantly decreased. (D) NAPA-loaded nanoparticles showed a concentration- dependent effect on different types of cancer cells, with a more prominent effect on cancer stem cells (CSCs).

Article Snippet: The fourth cell variant is patient-derived xenograft pancreatic cancer stem cells (CSCs) obtained from Celprogen.

Techniques: Blocking Assay, In Vitro, Encapsulation, Activity Assay, Incubation, Standard Deviation, Concentration Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Clinical significance of PNO1 as a novel biomarker and therapeutic target of hepatocellular carcinoma

doi: 10.1111/jcmm.18295

Figure Lengend Snippet: Effects of PNO1 knockout on the expression of genes involved in stemness, cell cycle, and inflammation in liver cancer cells. (A) Gene expression in Hep3B cells. Liver cancer Hep3B cells expressing either non‐targeting control (NTC) or PNO1 CRISPR/Cas9 were seeded. After 24 h, cells were harvested, and RNA was extracted to measure the expression of CD44, CCND1, p21, PTGS‐2, IL‐1α, IL‐8 and CXC‐8 by qRT‐PCR. Data represent mean ± SD. * = significantly different from control, p < 0.05. (B) Gene expression in HepG2 cells. Liver cancer HepG2 cells expressing either non‐targeting control (NTC) or PNO1 CRISPR/Cas9 were seeded. After 24 h, cells were harvested, and RNA was extracted to measure the expression of CD44, CCND1, p21, PTGS‐2, IL‐1α, IL‐8 and CXC‐8 by qRT‐PCR. Data represent mean ± SD. * = significantly different from control, p < 0.05.

Article Snippet: CD44 + /CD133 + human cancer stem cells (CSCs) were isolated from primary hepatocellular carcinoma (obtained from Celprogen, Torrance, CA) and grown in a well‐defined stem cell culture medium, as per the supplier's instructions.

Techniques: Knock-Out, Expressing, CRISPR, Quantitative RT-PCR

Journal: Journal of Cellular and Molecular Medicine

Article Title: Clinical significance of PNO1 as a novel biomarker and therapeutic target of hepatocellular carcinoma

doi: 10.1111/jcmm.18295

Figure Lengend Snippet: Effects of PNO1 knockout on cancer stem cell (CSC) characteristics. (A) PNO1 expression. CSCs were transduced with lentiviral particles expressing either non‐targeting control (NTC) or PNO1 CRISPR/Cas9, and the expression of PNO1 gene and protein was measured by qRT‐PCR and western blot analysis, respectively. Data represent mean ± SD. * = significantly different from control, p < 0.05. (B) % Colony formation. CSCs/NTC and CSCs/PNO1 CRISPR/Cas9 were seeded on dishes containing Matrigel. Number of colonies formed at 21 days were counted. Data represent mean ( n = 4) ± SD. * = significantly different from NTC, p < 0.05. (C) % Cell viability in spheroids. CSCs/NTC and CSCs/PNO1 CRISPR/Cas9 were seeded on ultralow attachment plate in suspension for 7 days to obtain primary spheroids. At the end of incubation period, spheroids were collected and reseeded for another week to obtain secondary spheroids. At the end of second incubation period (2 weeks), spheroids were collected and reseeded for another week to obtain tertiary spheroids. Cell viability in spheroids was measured by trypan blue assay at the end of 7, and 14, 21 days. Data represent mean ± SD. *, #, and @ = significantly different from control, p < 0.05. (D) Expression of CD44, CD133 and AFP by qRT‐PCR. CSCs expressing either NTC or PNO1 CRISPR/Cas9 were seeded. After 24 h, cells were harvested, and RNA was extracted to measure the expression of CD44, CD133 and AFP by qRT‐PCR. Data represent mean ± SD. * = significantly different from control, p < 0.05. (E) Expression of CD44, CD133 and AFP by immunohistochemistry. CSCs expressing either NTC or PNO1 CRISPR/Cas9 were seeded. After 24 hrs, CSCs were fixed and the expression of CD44, CD133 and AFP was measured by immunohistochemistry. Green colour = CD44, CD133 or AFP. Blue colour = nuclei. (F) Expression of pluripotency‐maintaining factors (cMyc, KLF4, Sox2, Oct4 and Nanog). CSCs expressing either NTC or PNO1 CRISPR/Cas9 were seeded. After 24 h, CSCs were harvested, and RNA was extracted to measure the expression of cMyc, KLF4, Sox2, Oct4 and Nanog. Data represent mean ± SD. * = significantly different from control, p < 0.05.

Article Snippet: CD44 + /CD133 + human cancer stem cells (CSCs) were isolated from primary hepatocellular carcinoma (obtained from Celprogen, Torrance, CA) and grown in a well‐defined stem cell culture medium, as per the supplier's instructions.

Techniques: Knock-Out, Expressing, Transduction, CRISPR, Quantitative RT-PCR, Western Blot, Suspension, Incubation, Immunohistochemistry

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A), Human pancreatic CSCs were grown in six-well ultralow attachment plates (Corning Inc., Corning, NY) at a density of 1,000 cells/ml in Celprogen pancreatic CSC medium at 37°C in a humidified atmosphere of 95% air and 5% CO 2 and treated with resveratrol (0–30 µM) for 7 days to obtain primary spheroids. At the end of incubation period, spheroids were collected, reseeded and treated with resveratrol for another week to obtain secondary spheroids. (B) Cell viability in spheroids was measured by trypan blue assay at the end of 7 and 14 days from above experiment. Data represent mean ± SD. *, &, #, % and @ = significantly different from controls, P<0.05. (C), Pancreatic CSCs were seeded in soft agar and treated with resveratrol (0–30 µM) for 21 days. At the end of incubation period, numbers of colonies were counted. Data represent mean ± SD. *, % and & = significantly different from control, P<0.05.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Incubation

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A–H), Pancreatic cancer cells were isolated from primary tumors and analyzed by flow cytometry using antibody against CD44, CD24, ESA, CD133, Oct4, and ALDH. (I), Expression of stem cell markers. RNA was isolated from normal pancreatic tissues, primary pancreatic cancer and pancreatic CSCs and the expression of CD24, CD133, CD44, ESA, Nanog, Notch1, MDR1 and ABCG2 was measured by q-RT-PCR. Data represent mean ± SD. *, ** @ = significantly different from controls, P<0.05.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Isolation, Flow Cytometry, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A), NOD/SCID mice were sc injected with 50 CD133 − CD44 − CD24 − ESA − cells on right flank and CD133 + CD44 + CD24 + ESA + CSCs on left flanks and tumor growth was observed for 20 days. (B), Pancreatic CSCs are similar to human primary tumors. Tumor sections from human primary pancreatic tumors, and pancreatic CSCs grown in NOD/SCID mice were stained with H&E, Statifin and S100P.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Injection, Staining

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A), Human pancreatic CSCs were grown in six-well ultralow attachment plates (Corning Inc., Corning, NY) at a density of 1,000 cells/ml in Celprogen pancreatic CSC medium at 37°C in a humidified atmosphere of 95% air and 5% CO 2 and treated with resveratrol (0–30 µM) for 7 days to obtain primary spheroids. At the end of incubation period, spheroids were collected, reseeded and treated with resveratrol for another week to obtain secondary spheroids. (B) Cell viability in spheroids was measured by trypan blue assay at the end of 7 and 14 days from above experiment. Data represent mean ± SD. *, &, #, % and @ = significantly different from controls, P<0.05. (C), Pancreatic CSCs were seeded in soft agar and treated with resveratrol (0–30 µM) for 21 days. At the end of incubation period, numbers of colonies were counted. Data represent mean ± SD. *, % and & = significantly different from control, P<0.05.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Incubation

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A and B), Activation of caspase-3/-7 and induction of apoptosis. Pancreatic CSCs were treated with resveratrol (0–30 µM) for 48 h, and caspase-3/-7 activity and apoptosis were measured by colorometric and TUNEL assay, respectively. Data represent mean ± SD. *, & and # = significantly different from control, P<0.05. (C and D), Pancreatic CSCs were treated with resveratrol (0–20 µM) for 48 h, and western blot analysis was performed to measure the expression of XIAP, Bcl-2, caspase-3 and cyclin D1. GAPDH was used as a loading control.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Activation Assay, Activity Assay, TUNEL Assay, Western Blot, Expressing

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A), Pancreatic CSCs were treated with resveratrol (0–20 µM) for 24 h. The expression of Nanog, Sox-2 and cMyc was measured by qRT-PCR. Data represent mean ± SD. * and @ = significantly different from control, P<0.05. (B), Pancreatic CSCs were treated with resveratrol (0–30 µM) for 48 h. The expression of Nanog, Oct-4 and Sox-2 was measured by the Western blot analysis. GAPDH was used as a loading control. (C), Inhibition of Nanog transcription by resveratrol. Pancreatic CSCs were transduced with Nanog reporter construct. Transduced cells were treated with resveratrol to examine the Nanog transcriptional activity. Data represent mean ± SD. *, #, @, & and ** = significantly different from control, P<0.05. (D), Pancreatic CSCs were treated with resveratrol (0–30 µM) for 36 h. The expression of ABCG2 was measured by qRT-PCR. Data represent mean ± SD. * and # = significantly different from control, P<0.05.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Inhibition, Transduction, Construct, Activity Assay

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A), Expression of Nanog. Pancreatic CSCs were transduced with scrambled or Nanog shRNA, and Western blot analyses were performed to examine the expression of Nanog and GAPDH. (B), Transduced cells were treated with resveratrol (0–30 µM) and spheres in suspension were grown for one week. Spheres were harvested, resuspended and cell viability was determined by trypan blue assay.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Expressing, Transduction, shRNA, Western Blot

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: Pancreatic CSCs were treated with resveratrol (0–0 µM) for 24 h. The expression of Zeb-1 (A), Snail (B) and Slug (c) was measured by the qRT-PCR. Data represent the mean ± S.D. * or # = significantly different from respective controls, P<0.05. (D), Invasion assay. Pancreatic CSCs were plated onto the Matrigel-coated membrane in the top chamber of the transwell and treated with resveratrol (0–30 µM) for 48 h. Cells invaded to the lower chambered were fixed with methanol, stained with crystal violet and counted. Data represent mean ± SD. *, & or # = significantly different from control, P<0.05. (E) Migration assay. Pancreatic CSCs were plated in the top chamber of the transwell and treated with resveratrol (0–30 µM) for 24 h. Cells migrated to the lower chambered were fixed with methanol, stained with crystal violet and counted. Data represent mean ± SD. *, & or # = significantly different from control, P<0.05.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Invasion Assay, Staining, Migration

Journal: PLoS ONE

Article Title: Resveratrol Inhibits Pancreatic Cancer Stem Cell Characteristics in Human and Kras G12D Transgenic Mice by Inhibiting Pluripotency Maintaining Factors and Epithelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0016530

Figure Lengend Snippet: (A), Pancreatic CSCs were isolated from Kras G12D mice (about 12 months old) using CD133, CD44, CD24, and ESA antibodies. The expression of these markers and Nanog and Oct-4 was confirmed by qRT-PCR. Data represent mean ± SD. * = significantly different from the same gene of Pdx-Cre mice at P<0.05. (B), CSCs were grown in suspension and treated with resveratrol (0–30 µM) for 7 days to obtain primary spheroids. At the end of incubation period, spheroids were collected, resuspended and treated with resveratrol for another week to obtain secondary spheroids. Cell viability in spheroids was measured by trypan blue assay at the end of 7 and 14 days. Data represent mean ± SD. *, %, &, ** and ## = significantly different from respective controls, P<0.05.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were cultured in either Celprogen's pancreatic CSC medium or DMEM supplemented with 1% N2 Supplement (Invitrogen), 2% B27 Supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen) and 1% antibiotic-antimycotic (Invitrogen) at 37°C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: Isolation, Expressing, Quantitative RT-PCR, Incubation