human pancreatic cancer stem cells cscs  (Celprogen Inc)

Journal: Journal of Cellular and Molecular Medicine

Article Title: EN2 Regulates Pancreatic Cancer Initiation, Progression, and Epithelial‐Mesenchymal Transition Through the Notch Signalling Pathway

doi: 10.1111/jcmm.71158

Figure Lengend Snippet: The expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. (A), Protein expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. Crude proteins were isolated, and EN2 expression was measured by Western blot analysis. β‐Actin was used as a loading control. (B), Expression of EN2 mRNA in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. RNA was isolated, and EN2 expression was measured by q‐RT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from HPNE ( p < 0.05). (C), Expression of EN2. Immunocytochemistry was performed to examine EN2 expression in HPNE, PANC‐1, and AsPC‐1 cells.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were obtained from Celprogen and cultured in a well‐defined medium according to the manufacturer's instructions.

Techniques: Expressing, Isolation, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Immunocytochemistry

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