150 mg/kg 5-fluorouracil (APP Pharmaceuticals) was injected i.v. into six- to ten-week old C57BL/6 SJL mice as previously described (14 (link)). Five days post-treatment, the bone marrow was harvested and cultured overnight with the following growth factors: stem cell factor (SCF; 100 ng/mL), IL-3 (6 ng/mL), IL-6 (10 ng/mL; all from Peprotech), and 5% (vol/vol) conditioned media from WeHi-3 cells (ATCC). 5 × 105 HSCs were injected i.v. into eight week-old C57BL/6 mice that had been lethally irradiated (900 rad) on the same day.
Wehi 3 cells
Manufactured by American Type Culture Collection
Sourced in China, Japan, United States
WeHi-3 cells are a murine myeloma cell line derived from a Balb/c mouse. They are known for their ability to secrete immunoglobulin and can be used as a source of monoclonal antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
B16f10 cells, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (1 mentions)
Real time pcr kit, by Takara Bio (1 mentions)
Atp detection assay kit, by Cayman Chemical (1 mentions)
Apyrase, by Merck Group (1 mentions)
Alexa fluor 546 conjugated anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
Anti gfp antibody, by Novus Biologicals (1 mentions)
Sodium monoiodoacetate mia, by Merck Group (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
4 protocols using wehi 3 cells
1
Hematopoietic Stem Cell Transplantation
2
Murine Cell Line Culturing Protocol
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The 6-8-week-old male BALB/c mice, C57BL/6 and 5-week-old male nude mice were provided by the Center for Laboratory Animal Sciences of Southern Medical University (Guangzhou, China). FDC-P1 cells (ATCC CRL-12103) and WEHI-3 cells (ATCC TIB-68) (Peking Union Medical University, Beijing, China) were maintained in Dulbecco’s modified Eagle’s medium (DMEM. Hyclone Ltd, Logan, UT) containing 10 % fetal bovine serum (FBS. Hyclone). For culture of FDC-P1 cells, 10 % of WEHI-3 cell-conditioned medium (WEHI-3/CM) was supplemented. CTLL-2 cells (ATCC TIB-214), murine A1.1 T cell hybridoma [35 (link)] (kindly gift from Dr. Yufang Shi, The Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai, China), B16F10 cells (ATCC CRL-6475), B16-GMCSF (B16F10 cells stably transfected with mouse gm-csf gene), DC2.4 and RAW264.7 cells were conserved in our laboratory and maintained in RPMI-1640 medium (Hyclone) supplemented with 10 % FBS. All of the five cell lines were derived from C57BL/6 mouse.
3
Mast Cell Tryptase and PAR2 Signaling
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The following commercial materials were used: Alexa Fluor 546 conjugated anti-rabbit IgG antibody and Alexa Fluor 488-conjugated anti-goat IgG antibodies were from Thermo Fisher Scientific (Yokohama, Japan); anti-mast cell tryptase (V-13) antibody against mMCP-6 was from Santa Cruz Biotechnology (Santa Cruz, CA); anti-PAR2 antibody was from Bioss Inc. (Woburn, MA); anti-NF-H antibody was from Gene Tex Inc. (Irvine, CA); anti-GFP antibody was from Novus Biologicals (Centennial, CO); PAR2 antagonist peptide (FSLLRY-NH2) (TOCRIS), PAR2 agonist peptide (SLIGKV-NH2) (TOCRIS) and ATP Detection Assay Kit (Cayman Chemicals) were from Funakoshi Co., Ltd. (Tokyo, Japan); WEHI-3 cells were from American Type Culture Collection; RNA extraction kit (Thermo Fisher Scientific) and real time PCR kit (TAKARA) were from Takara Bio Inc. (Kusatsu, Japan); tryptase substrate (S-2288) was from Chromogenix (Milano, Italy); ATP, sodium monoiodoacetate (MIA) and apyrase were from Sigma-Aldrich Japan (Tokyo, Japan).
4
Culturing Murine Myelomonocytic Leukemic Cells
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WEHI-3 cells, a murine myelomonocytic leukemic cell line, were obtained from the American Type Culture Collection (Rockville, MD, USA). These cells were grown in RPMI-1640 supplemented with 10% heat-inactivated FBS, 2 mM glutamine, and 1% penicillin-streptomycin-neomycin in a 5% CO2 humidified incubator at 37°C. Cultures were harvested and cell numbers enumerated by hemocytometer analysis of cell suspensions.
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