Bulletkit

Manufactured by Lonza

Sourced in United States, Switzerland, Belgium, Germany

The BulletKit is a compact and versatile laboratory equipment designed for various scientific applications. It serves as a multi-purpose tool, offering essential functionalities for researchers and laboratory professionals. The core function of the BulletKit is to facilitate sample preparation, processing, and analysis tasks in a streamlined and efficient manner. The product specifications and capabilities are presented in a factual and unbiased manner, without extrapolation on the intended use.

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Lab products found in correlation

138 protocols using bulletkit

Adipogenic and Osteogenic Differentiation of hADSCs

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In order to evaluate the differentiation potency of purified hADSCs into adipocytes and osteocytes, we cultured 2nd passage of hADSCs in human mesenchymal stem cell adipogenic differentiation medium BulletKit (Cat# PT-3004, Lonza, Walkersville, MD, USA) or osteogenic differentiation medium BulletKit (Cat# PT-3002, Lonza). After 2–3 weeks of culture, adipogenic differentiation was identified using AdipoRed (Cat# PT-7009, Lonza, Walkersville, MD) staining for the lipid droplets and osteogenic differentiation was characterized using Alizarin Red (Cat# ARD-A1, PG Research, Kodaira, Tokyo, Japan) staining for the mineralized matrix. As control cells, human ADSCs were purchased from Lonza (Cat# PT-5006) and human dermal fibroblasts (Cat# KF-4109) were obtained from Kurabo, Chuo-ku, Osaka, Japan.

Nomura M., George J., Hashizume C., Saito T., Ueda Y., Ishigaki Y., Tsuchishima M, & Tsutsumi M. (2022). Surgical implantation of human adipose derived stem cells attenuates experimentally induced hepatic fibrosis in rats. Molecular Medicine, 28, 143.

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Fibroblast Cultures for Mitochondrial Disease

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Primary skin fibroblasts from two mitochondrial disease patients and age-matched healthy controls were cultured in a fibroblast growth medium (FGM; Lonza) constituting fibroblast basal medium (FBM; Lonza) supplemented with gentamicin/amphotericin B (antibiotic/antifungal) and growth factors (rhFGF-B, insulin, fetal bovine serum; all from BulletKits®, Lonza Cat. No. CC-3132 FGM™-2 BulletKit™). Cells were transfected with RNAiMax (Invitrogen) according to the manufacturer’s protocol. The small interfering RNA (siRNA) used was siDrp1, as described previously [30 (link)].

Tokuyama T., Hirai A., Shiiba I., Ito N., Matsuno K., Takeda K., Saito K., Mii K., Matsushita N., Fukuda T., Inatome R, & Yanagi S. (2020). Mitochondrial Dynamics Regulation in Skin Fibroblasts from Mitochondrial Disease Patients. Biomolecules, 10(3), 0.

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Prostate Cancer Cell Line Characterization

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Human PCA PC3, DU-145, LNCaP and 22Rv1 cells were from ATCC. C4-2B cells were from ViroMed Laboratories. PCA cell lines were tested and authenticated by DNA profiling for polymorphic short tandem repeat (STR) markers at University of Colorado Cancer Center DNA Sequencing & Analysis Core. RPMI 1640 media, other cell culture materials, TGFβ1 ELISA kit, and CAS-Block were from Invitrogen. PrSCs, SCGM, and Bullet-kits were from Lonza. Prostate CAFs were from a prostatectomy specimen removed at Wake Forest University [13 (link)]. Pathology of specimen was verified by two board-certified pathologists (AC and JS). No patient identifiers were retained and use of discarded tissue was not considered human subjects research by Wake Forest University IRB. DAPI and silibinin were from Sigma, IL-6 from Cell Signaling, and TGFβ1, TGFβ2, and goat IgG isotype control antibody were obtained from Gibco. TGFβ2 ELISA kit and TGFβ2-neutralizing antibody were obtained from R&D, antibodies to IL-6, α-SMA and FAP (fibroblast activation protein) from Abcam, antibody to vimentin and HRP conjugated streptavidin from Santa Cruz, 3,3′-diaminobenzidine (DAB) peroxidase substrate kit from Vector Labs, biotinylated antibodies and mouse IgG's from DAKO, and transwell invasion chambers from BD Biosciences.

Ting H., Deep G., Jain A.K., Cimic A., Sirintrapun J., Romero L.M., Cramer S.D., Agarwal C, & Agarwal R. (2014). Silibinin Prevents Prostate Cancer Cell-Mediated Differentiation of Naïve Fibroblasts into Cancer-Associated Fibroblast Phenotype by Targeting TGF β2. Molecular carcinogenesis, 54(9), 730-741.

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Culturing Normal Human Astrocytes and Glioblastoma Cell Lines

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Normal human astrocytes (NHAs) were obtained from Lonza (Basel, Switzerland) and cultured in astrocyte growth media (Lonza) containing 3% fetal bovine serum (FBS), 4.5% glucose and reagents from BulletKits (Lonza) in a humidified incubator at 37°C and 5% CO₂. Human GBM cell lines U87-MG, U251 and HEK-293T were purchased from Chinese Academy of Sciences Cell Bank (Shanghai, People’s Republic of China), and were cultured in DMEM (Hyclone, Logan, UT, USA) supplemented with 100 units of penicillin/mL, 100 ng of streptomycin/mL as well as 10% FBS. All the cells were maintained at 37°C in a humidified atmosphere with 5% CO₂.

Zhang C., Yang M., Li Y., Tang S, & Sun X. (2018). FOXA1 is upregulated in glioma and promotes proliferation as well as cell cycle through regulation of cyclin D1 expression. Cancer Management and Research, 10, 3283-3293.

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Differentiation of hMSCs into Adipocytes and Osteocytes

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Untransduced hMSCs as well as CEETnl2- and CEETnl2Is2 transduced hMSC were differentiated to adipocytes and osteocytes as previously described76 (link). Briefly, hMSCs were plated in 6-well plates at a density of 20,000 cells/cm² for adipogenesis, 10,000 cells/cm² for osteogenesis and incubated in the adipogenic and osteogenic MSCs differentiation BulletKits respectively (lonza, Basel, Switzerland).

Benabdellah K., Muñoz P., Cobo M., Gutierrez-Guerrero A., Sánchez-Hernández S., Garcia-Perez A., Anderson P., Carrillo-Gálvez A.B., Toscano M.G, & Martin F. (2016). Lent-On-Plus Lentiviral vectors for conditional expression in human stem cells. Scientific Reports, 6, 37289.

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Culturing Skin Cells for Research

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NHEKs obtained from neonatal foreskin were purchased from Lonza (Basel, Switzerland) and cultured in keratinocyte growth medium (KBM Gold) with BulletKit (Lonza, Walkersville, MD, USA) containing insulin, human epidermal growth factor, bovine pituitary extract, hydrocortisone, epinephrine, transferrin, and gentamicin/amphotericin B. The cells were serially passaged until 70–80% confluence was achieved, which was no more than three times. Cell proliferation and cytotoxicity were measured with a Cell Counting Kit-8 (Dojindo, Kumamoto, Japan). Moderately pigmented human epidermal melanocytes were purchased from Cascade Biologics (Portland, OR, USA) and maintained and passaged in Medium 254 (#M254500) supplemented with Human Melanocyte Growth Supplement (Life Technologies, Carlsbad, CA, USA), 10% fetal bovine serum, 100 U/mL of penicillin G, and 100 μg/mL of streptomycin sulfate. The human epidermal melanocytes were incubated at 37 °C with 5% CO₂ and regularly passaged at a density of 80% (1:8 ratio). Cell viability was measured using the Cell Counting Kit-8 (Dojindo, Kumamoto, Japan).

Kim J., Lee J, & Choi H. (2021). Intense Pulsed Light Attenuates UV-Induced Hyperimmune Response and Pigmentation in Human Skin Cells. International Journal of Molecular Sciences, 22(6), 3173.

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Culturing Cos-7 and HUVEC Cells

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Cos-7 cells (ATCC, CRL-1651) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Corning, 10013CV) containing L-glutamine and sodium pyruvate and supplemented with 10% fetal bovine serum (Gibco, 10438-026) and 100 IU/ml penicillin-streptomycin (Gibco, 15070-063). Human Umbilical Vein Endothelial Cells (Pooled HUVECs, Lonza, C2519A) were cultured in Endothelial Cell Growth Medium-2 (EGM2, Lonza, CC-3156) supplemented with the BulletKit (Lonza, CC-3162). Cells were maintained at 37 °C and 5% CO₂. Cells were passaged at 90% confluency, resuspended in fresh media, and one-fifth was plated on 100 mm Tissue Cell Culture Dishes (Falcon, 353003). For HUVECs full EGM-2 media was first placed onto a cell culture dish and left to equilibrate into the incubator for 30 min.

Nawara T.J., Williams YD I.I., Rao T.C., Hu Y., Sztul E., Salaita K, & Mattheyses A.L. (2022). Imaging vesicle formation dynamics supports the flexible model of clathrin-mediated endocytosis. Nature Communications, 13, 1732.

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Cell Culture of Epithelial and Endothelial Cells

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Bronchial Epithelial cell line BEAS-2B and human epithelial carcinoma cell-line NCI H-1299 was procured from ATCC (Middlesex, UK). BEAS-2B was cultured in BEGM media supplemented with bullet kit from Lonza, THP-1 and H-1299 cells were maintained in RPMI 1640 supplemented with 10% FCS that was depleted of FBS exosomes by spinning it at 100,000 g for 12 hrs. HUVECs were isolated from human umbilical cord and were cultured in M199 media supplemented with ECGF (Sigma, USA). Experiments with Human umbilical cords were performed as per guidelines and protocols approved by the CSIR-IGIB Human Ethics Committee. Prior informed consent from volunteers was obtained for the collection of material for this study.

Kulshreshtha A., Singh S., Ahmad M., Khanna K., Ahmad T., Agrawal A, & Ghosh B. (2019). Simvastatin mediates inhibition of exosome synthesis, localization and secretion via multicomponent interventions. Scientific Reports, 9, 16373.

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Endothelial Cell Isolation and Analysis

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GSCs were grown in endothelial cell growth medium (EGM)-2 supplemented with the bullet kit (Lonza) for 3 days at 37°C. Cells were dissociated, spun, counted, re-suspended in FACS buffer (2% inactivated fetal calf serum in PBS), incubated with FITC-conjugated anti-human CD31 (Biolegend) for 30 min at room temperature, washed, fixed in 4% paraformaldehyde, washed, re-suspended in 2% FACS buffer and sorted by LSRII flow cytometer (BD Biosciences). Data were analyzed with FlowJo software v.10.1 (Tree Star).

Saha D., Wakimoto H., Peters C., Antoszczyk S.J., Rabkin S.D, & Martuza R.L. (2018). Combinatorial effects of VEGFR kinase inhibitor axitinib and oncolytic virotherapy in mouse and human glioblastoma stem-like cell models. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(14), 3409-3422.

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Culturing Melanoma, Glioblastoma, and Endothelial Cells

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The melanoma cell line, Yumel 0106 (homozygous for the BRAF V600E mutation), kindly provided by Xandra O. Breakefield, was cultured in OptiMem (Invitrogen) containing 10% fetal bovine serum (FBS) and 5% penicillin/streptomycin. The GBM cell line DBTRG-05MG (heterozygous for the BRAF V600E mutation, ATCC CRL-2020) was cultured in RPMI-1640 (ThermoFisher) containing 10% FBS and 5% penicillin/streptomycin. Human brain microvascular endothelial cells (HBMVEC, BRAF WT) were kindly provided by Xandra O. Breakefield and cultured in Microvascular Endothelial Cell Growth Medium-2 (BulletKit, Lonza) containing 10% FBS and 5% penicillin/streptomycin. gDNA and mRNA was isolated from cultures at 50–70% confluency and all cell lines are negative for mycoplasma contamination (Mycoplasma PCR Detection Kit; Applied Biological Materials).

Kang K.M., Muralidharan K., Yekula A., Small J.L., Rosh Z.S., Jones P.S., Carter B.S, & Balaj L. (2021). Blood-Based Detection of BRAF V600E in Gliomas and Brain Tumor Metastasis. Cancers, 13(6), 1227.

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