Journal: iScience

Article Title: Feasibility and barriers to rapid establishment of patient-derived primary osteosarcoma cell lines in clinical management

doi: 10.1016/j.isci.2024.110251

Figure Lengend Snippet: Studies that established PCCL from primary resected tumor

Article Snippet: Thanindratarn et al. , OSA1777 , 1. 19, F 2. Left proximal femur 3. Not specified , 1. Tumor tissues directly harvested in the operating theater. 2. Placed in sterile normal saline within a sealed microcentrifuge tube on ice for immediate transport to the laboratory , 1. Specimens were washed three times with RPMI 1640 containing 1% penicillin/streptomycin (100 U/mL. Minced with a razor blade 2. Not specified , 1. Cultured in RPMI-1640 medium supplemented with 10% FBS and 1% penicillin/streptomycin, in tissue culture flasks. 2. Standard conditions at 37°C in an atmosphere of 95% air:5% CO2 3. Media replaced every 3–4 days 4. Harvested using 0.25% trypsinization at 80–90% confluence, with subculture into new flasks. >50 passages 5. 6 months , 1. a) STR profile >unique genomic identity without matching ATCC, DSMZ repositories >AMEL locus of X chromosome consistent with female patient origin >no contamination by another human cell line detected >no contamination by mouse DNA detected 1. b) Western blot >EMT markers (vimentin+/cytokeratin-) >tumor markers (p53-/MDM2+) 2. Morphology >spindle-shaped with a round to oval nuclei Doubling time >60h Chemosensitivity assay >in 2D culture: Cell viability decreased after 5days exposure. More sensitive to triple treatment (MAP) compared with single treatment >in 3D culture: decreased spheroid size after 7days exposure Spheroid formation assay >cluster formation in 3D culture by day 5 >similar growth rate to MNNG-HOS and U2OS 3. Not specified.

Techniques: Incubation, Sterility, Saline, Cell Culture, Western Blot, Tube Formation Assay, Injection, Staining, Invasion Assay, MTT Assay, Immunohistochemistry

Journal: STAR Protocols

Article Title: Mouse intraductal modeling of primary ductal carcinoma in situ

doi: 10.1016/j.xpro.2023.102526

Figure Lengend Snippet: Intraductal injection setup (A) camera. (B) heating pad. (C) 5 mL syringe connected to isoflurane tube. (D) Hamilton syringe. (E) Dumont tweezer. (F) 15 mL Falcon tube with PBS. (G) Ice bucket with samples and stock of Evans Blue. (H) pipet tips. (I) screen. (J) Eppendorf tubes. (K) 20 μL pipet. (L) 70% ethanol. (M) razor blade. (N) 5 mL syringe connected to isoflurane on a standard for shaving. (O) anesthesia induction box. (P) Cage with mice.

Article Snippet: Timing: 1 day Ensure you have all the equipment necessary for intraductal injections (tissues, razor blades, 70% Ethanol, PBS, 15 mL Falcon tube, 20 μL pipet and tips, 1.5 mL Eppendorf tubes, anesthesia induction box, waste box, Evans blue (2 mg/mL), syringes, instruments (microscope (EVO CAM II with TV screen), tweezers, Hamilton syringe) and a flow cabinet with isoflurane.

Techniques: Injection

Journal: STAR Protocols

Article Title: Mouse intraductal modeling of primary ductal carcinoma in situ

doi: 10.1016/j.xpro.2023.102526

Figure Lengend Snippet: Impression of intraductal injection Also see .

Article Snippet: Timing: 1 day Ensure you have all the equipment necessary for intraductal injections (tissues, razor blades, 70% Ethanol, PBS, 15 mL Falcon tube, 20 μL pipet and tips, 1.5 mL Eppendorf tubes, anesthesia induction box, waste box, Evans blue (2 mg/mL), syringes, instruments (microscope (EVO CAM II with TV screen), tweezers, Hamilton syringe) and a flow cabinet with isoflurane.

Techniques: Injection

Journal: STAR Protocols

Article Title: Mouse intraductal modeling of primary ductal carcinoma in situ

doi: 10.1016/j.xpro.2023.102526

Figure Lengend Snippet: Success intraductal injection (A) Successful intraductal injection. (B) Successful intraductal injection as seen under the skin. (C) Unsuccessful intraductal injection.

Article Snippet: Timing: 1 day Ensure you have all the equipment necessary for intraductal injections (tissues, razor blades, 70% Ethanol, PBS, 15 mL Falcon tube, 20 μL pipet and tips, 1.5 mL Eppendorf tubes, anesthesia induction box, waste box, Evans blue (2 mg/mL), syringes, instruments (microscope (EVO CAM II with TV screen), tweezers, Hamilton syringe) and a flow cabinet with isoflurane.

Techniques: Injection