X vivo

Manufactured by Lonza

Sourced in Switzerland, Belgium, Germany, United States, United Kingdom

X-VIVO is a serum-free, xeno-free cell culture medium designed for the expansion and maintenance of a variety of cell types, including T cells, NK cells, and stem cells. It is formulated to support optimal cell growth and function without the use of animal-derived components.

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Lab products found in correlation

Interleukin 2 (il 2), by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Dynabeads human t activator cd3 cd28, by Thermo Fisher Scientific (3 mentions) Human ab serum, by Merck Group (3 mentions) Human serum, by Euroclone (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Automacs pro, by Miltenyi Biotec (2 mentions) Hil 3, by Miltenyi Biotec (2 mentions) H tpo, by Miltenyi Biotec (2 mentions) Fetal bovine serum (fbs), by Lonza (2 mentions) Facsaria 2, by BD (2 mentions) Sodium pyruvate, by Thermo Fisher Scientific (2 mentions) Rhil 2, by Thermo Fisher Scientific (2 mentions) Htgf β1, by R&D Systems (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Hm40 3, by BD (2 mentions) Il 15, by Thermo Fisher Scientific (2 mentions) Interleukin 2 (il 2), by Novartis (2 mentions) Dnase, by STEMCELL (2 mentions) Anti pe microbeads, by Miltenyi Biotec (2 mentions)

Close spelling variants of this product

X-VIVO 15 medium (408 citations) X-VIVO 15 (360 citations) X-VIVO 10 medium (60 citations) X-Vivo media (37 citations) X-VIVO 20 medium (33 citations) X-vivo 20 serum-free medium (8 citations) X-VIVO 20 media (8 citations) X-VIVO 15 serum-free cell-culture medium (6 citations) X-VIVO 15 chemically defined medium (3 citations) X-Vivo 15 cell medium (3 citations)

56 protocols using x vivo

Lentiviral Vector-Mediated Gene Transduction of T-Cells

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The bidirectional SIN.LV.PGK.BACH2 and SIN.LV.PGK.STAT5B plasmid constructs were generated using Gateway Technology (for details refer to Supplementary Methods). All concentrated VSV.G-pseudotyped LV stocks were produced by transient transfection of 293 T cells (CRL-3216, ATCC), as previously reported^{7 (link)}.
For transduction, Naïve T lymphocytes were pre-activated for 24 h in X-VIVO (Lonza), 5% human serum (Euroclone), in presence of anti-CD3/CD28 activating beads (Dynabeads Human T cell activator CD3/CD28, Life Technologies). Treg and Teff cells were pre-activated for 24 h in X-VIVO (Lonza), 5% human serum (Euroclone), in presence of Human Treg Expander beads (Life Technologies). Cells were infected with the indicated LV at MOI 20.

Cesana D., Santoni de Sio F.R., Rudilosso L., Gallina P., Calabria A., Beretta S., Merelli I., Bruzzesi E., Passerini L., Nozza S., Vicenzi E., Poli G., Gregori S., Tambussi G, & Montini E. (2017). HIV-1-mediated insertional activation of STAT5B and BACH2 trigger viral reservoir in T regulatory cells. Nature Communications, 8, 498.

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Isolation and Expansion of Human CD34+ Cells

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Mononuclear cells were isolated from umbilical cord blood from anonymous healthy donors by density centrifugation using Ficoll (Biocoll, Merck Millipore). Human CD34+ cells were then enriched in the sample by immunomagnetic beads using an AutoMACSpro (Miltenyi Biotec). After collection, enriched CD34+ cells were frozen in a cryopreservation medium containing 90% of fetal bovine serum (Eurobio) and 10% of dimethylsulfoxide (Sigma) and stored in liquid nitrogen.
After thawing, the CD34+ cells were cultured in a 96-well plate in a humidified 5% CO2 incubator at 37°C. Cells were cultured in prestimulation medium made of XVivo (Lonza) supplemented with penicillin/streptomycin (respectively, 100 U/mL and 100 μg/mL; Gibco, Thermo Fisher Scientific), 50 ng/ml h-FLT3-ligand, 25 ng/ml h-SCF, 25 ng/ml h-TPO, and 10 ng/ml h-IL3 (Miltenyi) final concentration.

Parmentier R., Racine L., Moussy A., Chantalat S., Sudharshan R., Papili Gao N., Stockholm D., Corre G., Fourel G., Deleuze J.F., Gunawan R, & Paldi A. (2022). Global genome decompaction leads to stochastic activation of gene expression as a first step toward fate commitment in human hematopoietic cells. PLOS Biology, 20(10), e3001849.

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Cellular Uptake Assay with pHrodo

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Experimental procedure was performed according to manufacturer's instructions. In brief, cells were washed and growth medium was replaced with X‐Vivo (Lonza). pHrodo™ dextran was added to the cells at a final concentration of 20–100 μg/ml, and cells were incubated at 37°C for the indicated time points. Cells were washed with pre‐warmed, dye‐free medium at pH 7.4. Cells were returned to dye‐free medium at pH 7.4, and flow cytometric analysis was performed.

Khatamzas E., Hipp M.M., Gaughan D., Pichulik T., Leslie A., Fernandes R.A., Muraro D., Booth S., Zausmer K., Sun M., Kessler B., Rowland‐Jones S., Cerundolo V, & Simmons A. (2017). Snapin promotes HIV‐1 transmission from dendritic cells by dampening TLR8 signaling. The EMBO Journal, 36(20), 2998-3011.

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Transducing iPSC-Derived RPE Cells

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All RPE p2 cells were matured for 4 weeks on transwells before transduction. AAV2/Anc80 AAP.CASI.V5.PRPF31-mCHERRY.RBG was suspended in X-VIVO (BE04-743Q, Lonza, Basel, Switzerland) with Normocin (ant-nr-1, InvivoGen, San Diego, CA, USA) to achieve an MOI of 25,000, 50,000, and 100,000 vector genomes (vg)/cell in a final volume of 50 μL for low-density cultures or 100 μL for high-density cultures. AAV was added to the iPSC-RPE cells on transwells and incubated overnight. Media were changed the next day and twice/week afterward. All RPE cells were cultured for 4 weeks before performing a phagocytosis assay.

Brydon E.M., Bronstein R., Buskin A., Lako M., Pierce E.A, & Fernandez-Godino R. (2019). AAV-Mediated Gene Augmentation Therapy Restores Critical Functions in Mutant PRPF31+/− iPSC-Derived RPE Cells. Molecular Therapy. Methods & Clinical Development, 15, 392-402.

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Culturing Human Fetal Astrocytes and Monocyte-Derived Macrophages

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Human fetal astrocytes (HFAs) (ScienCell Research Laboratories), isolated from second trimester fetal cerebral cortex, were cultured in astrocyte media (ScienCell Research Laboratories) with 1% penicillin and streptomycin (P/S), 2% fetal bovine serum (FBS), and 1% astrocyte growth supplement (optimized insulin, transferrin, fibroblast growth factor, insulin-like growth factor 1, hydrocortisone, and progesterone). Blood-derived monocytes were isolated from peripheral blood mononuclear cells using Human Monocyte Isolation Kit II (Miltenyi Biotec) according to manufacturer’s instructions. Monocytes were cultured in X-vivo (Lonza) with 1% heterologous human serum (Sigma) for 7 to 10 days, during which time they differentiated into monocyte-derived macrophages (MDMs). TZM-bl cells (NIH AIDS Research and Reference Reagent Program) and 293T cells were grown in DMEM (Sigma) with 1% P/S and 10% FBS.

Russell R.A., Chojnacki J., Jones D.M., Johnson E., Do T., Eggeling C., Padilla-Parra S, & Sattentau Q.J. (2017). Astrocytes Resist HIV-1 Fusion but Engulf Infected Macrophage Material. Cell Reports, 18(6), 1473-1483.

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Induction of Human Regulatory T Cells

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Human PBMCs were obtained from blood samples of healthy donors (Shanghai Blood Center) with ethical approval from Shanghai Blood Center Ethics Committee. Human CD4⁺CD25^lowCD127^highCD45RA⁺ naïve CD4⁺ T cells were sorted by BD FACS Aria II cell sorter and differentiated into inducible T_reg cells in X-VIVO (Lonza, Malkersville, USA, Cat#04-418Q) medium supplemented with 10% fetal bovine serum (Invitrogen, Maltham, USA, Cat#10100147), 1% GlutaMAX, 1% sodium pyruvate, 1% minimum essential medium with nonessential amino acids, 1% penicillin-streptomycin, 100 U/mL IL2 (R&D systems, Minneapolis, USA, Cat#202-IL), and 5 ng/mL transforming growth factor-β (R&D Systems, Cat#240-B), in the presence of Dynabeads Human T-activator CD3/CD28 (Gibco, Cat#11 132D) at a bead-to-cell ratio of 1:4. Approximately 7 days later, the differentiation efficiency reached at least 90% and could be used for analysis.

Deng B., Yang B., Chen J., Wang S., Zhang W., Guo Y., Han Y., Li H., Dang Y., Yuan Y., Dai X., Zang Y., Li Y, & Li B. (2022). Gallic acid induces T-helper-1-like Treg cells and strengthens immune checkpoint blockade efficacy. Journal for Immunotherapy of Cancer, 10(7), e004037.

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Intestinal Tissue Cytokine Stimulation

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Immediately after collection, biopsies were cut in half following the intestinal layers’ orientation and split in two tubes containing a physiological saline solution. Once they reached the laboratory, the specimens were transferred to sterile tubes with culture medium (X-VIVO, Lonza, Verviers, Belgium) supplemented with 10% human AB serum (Sigma-Aldrich, Milano, Italy) and 2 mmol/L L-glutamine (EuroClone, Milano, Italy). The intestinal tissue was exposed to 250 ng/mL MDP (N-Acetylmuramyl-L-alanyl-D-isoglutamine hydrate, Sigma-Aldrich) or medium alone, in the presence or absence of antibiotics: 100 U/mL penicillin (EuroClone) and 0.1 mg/mL streptomycin (EuroClone).
After 24 h culture at 37 °C in a humidified atmosphere (95% air and 5% CO₂), supernatants were collected and stored at -20 °C for cytokine analysis.

Loganes C., Valencic E., Pin A., Marini E., Martelossi S., Naviglio S., De Leo L., Not T., Monasta L., Tommasini A, & Marcuzzi A. (2016). Ex vivo response to mucosal bacteria and muramyl dipeptide in inflammatory bowel disease. World Journal of Gastroenterology, 22(44), 9734-9743.

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Cultivation and Transduction of Leukemic Cell Lines

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Leukemic cell lines NALM-6 and BV173 were purchased from the American Type Culture Collection (ATCC) and cultured in RPMI 1640 (BioWhittaker) supplemented with 10% FBS (Lonza), 100 IU/mL penicillin/streptomycin, and glutamine. The ALL-CM cell line was provided by Fred Falkenburg (Leiden University Medical Center, Leiden, The Netherlands) and maintained in culture in X-VIVO (Lonza) with 3% human serum (Euroclone) and 100 IU/mL penicillin/streptomycin. For in vivo experiments, the NALM-6 cell line was transduced with a lentiviral vector encoding the secreted Lucia luciferase (Lucia⁺NGFR⁺ NALM-6), as previously reported (57 (link)).

Arcangeli S., Bove C., Mezzanotte C., Camisa B., Falcone L., Manfredi F., Bezzecchi E., El Khoury R., Norata R., Sanvito F., Ponzoni M., Greco B., Moresco M.A., Carrabba M.G., Ciceri F., Bonini C., Bondanza A, & Casucci M. (2022). CAR T cell manufacturing from naive/stem memory T lymphocytes enhances antitumor responses while curtailing cytokine release syndrome. The Journal of Clinical Investigation, 132(12), e150807.

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Derivation of MDS-like Cells from iPSCs

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TP53 wild-type and knock-out K562 cells have been described previously and were propagated in IMDM with 10% fetal calf serum [24] . Transfection of K562 cells was done with the SF cell line 4D-Nucleofector X Kit S from Lonza. Primary human HSPCs were cultured in XVIVO (Lonza) with SCF, FLT3 ligand, thrombopoietin (TPO), IL-3, and IL-6. Methylcellulose colony formation assays were performed using Methocult SF H4636 supplemented with FLT3-ligand, IL-6, and TPO. After 14 days, colonies were scored blindly and cells counted after diluting the cultures, and cells replated. In some experiments, single cell derived colonies were picked for multiplexed genotyping.
For derivation of KRAS G12D or FLT3-ITD-driven 'high-grade' MDS-like cells, retroviruses encoding these oncogenes were introduced either into the undifferentiated iPSCs or serially replating 'low-grade' MDS-like cells from colony assays to establish cultures of semi-transformed HPCs stably growing in suspension (high-grade MDS-like cells; MDS-high). MDS-high cells were cultured in SFEM (Stem Cell Technologies) with 100 ng/ml SCF, 50 ng/ml TPO, 100 ng/ml FLT3-ligand, 10 ng/ml IL-3, and 10 ng/ml IL-6 routinely and for downstream experiments. Both KRAS G12D and FLT3-ITD-driven lines were independently established at least three times using this methodology. All lines were tested for Mycoplasma every two weeks.

Marion W., Koppe T., Chen C.C., Wang D., Frenis K., Fierstein S., Sensharma P., Aumais O., Peters M., Ruiz-Torres S., Chihanga T., Boettcher S., Shimamura A., Bauer D.E., Schlaeger T., Wells S.I., Ebert B.L., Starczynowski D., da Rocha E.L, & Rowe R.G. (2023). RUNX1 mutations mitigate quiescence to promote transformation of hematopoietic progenitors in Fanconi anemia. Leukemia, 37(8).

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Generation of Induced Regulatory T Cells

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Human CD4⁺CD25loCD45RAhi naive T cells were flow sorted from the PBMCs of healthy donor using BD FACSAria II (BD Bioscience). Cells were differentiated into induced Treg (iTreg) using anti-CD3/CD28 DynaBeads at a 1:4 bead-to-cell ratio in cell culture media [X-VIVO (Cat# 04–418Q, Lonza), supplemented with 10% FBS (Cat# 10100147C, GIBCO), 1% L-Glutamine-100X (Cat# 335050061, GIBCO), 1% MEM Non-Essential Amino Acids-100X (Cat# 11140050, GIBCO), 1 mM sodium pyruvate (Cat# 11360070, GIBCO), 1% Antibiotic-Antimycotic 100x (Cat# 15240112, GIBCO)],
100 U/mL rhIL-2 (Cat# 200–02, Peprotech) and 5 ng/mL hTGF-β1 (Cat# 7754-BH-100/CF, R&D). After 5 days of culture, cells were characterized by intracellular flow cytometry for Foxp3 expression.

Ke S., Xie F., Guo Y., Chen J., Wang Z., Yu Y., Geng H., Xu D., Liu X., Xia X., Yu F., Zhu C., Zhang Z., Zhao G., Li B, & Zhao W. (2022). High-level of intratumoral GITR+ CD4 T cells associate with poor prognosis in gastric cancer. iScience, 25(12), 105529.

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