Lamina propria mononuclear cells (LPMCs) were isolated from intestinal resection samples as previously described.64 (link) The dead cell removal kit (#130-090-101, Miltenyi Biotec) was used to remove dead cells from LPMCs. The magnetic-activated cell sorting and fluorescence-activated cell sorting were used to obtain pure cell populations. CD4+ TRM cells were isolated from the LPMCs by negative selection using human CD4+ T-cell memory effector cell isolation kit23 (link) (#130-094-125, Miltenyi Biotec). In brief, the magnetically labeled B cells, naive T cells, antigen-presenting cells, CD8+ T cells, non-αβ+ T cells, and effector and central memory CD4+ T cells were positively selected. Then, CD4+ TRM cells in LPMCs were purified using fluorescence-activated cell sorter according to established surface markers CD69 and CD103 (FITC anti-human CD69 antibody, BioLegend 310904; PE/cyanine7 anti-human CD103 antibody, BioLegend 350212).22 (link) For the ex vivo culture of purified CD4+ TRM cells, complete X-VIVO 15 hematopoietic cell medium was obtained from Lonza (#BE02-060Q).
X vivo 15 hematopoietic cell medium
Manufactured by Lonza
Sourced in Switzerland
X-VIVO 15 is a serum-free, chemically defined medium designed for the culture and expansion of hematopoietic cells, including human and animal cells. It is optimized to support the growth and differentiation of hematopoietic progenitor cells.
Automatically generated - may contain errors
Lab products found in correlation
Fitc anti human cd69 antibody, by BioLegend (1 mentions)
Dead cell removal kit, by Miltenyi Biotec (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Cd27 fitc, by BD (1 mentions)
Cd3 percp, by BD (1 mentions)
Alamarblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
L glutamate, by Thermo Fisher Scientific (1 mentions)
Tryple express enzyme solution, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
2 protocols using x vivo 15 hematopoietic cell medium
1
Isolation and Purification of Intestinal CD4+ TRM Cells
2
Bladder Cancer Cell Lines Characterization
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Cell lines, reagents and chemicals. T24 human bladder carcinoma (BCRC No.60062) and BFTC905 human bladder papillary transitional cell carcinoma (BCRC No.60068) cell lines were purchased from Bioresources Collection and Research Center (Hsin Chu City, Taiwan, ROC). Ficoll ® Paque Plus was purchased from GE Healthcare Life Sciences (Parramatta NSW, Australia). Carboplatin was purchased from Accela ChemBio. Inc. (San Diego, CA, USA). X-VIVO 15 hematopoietic cell medium was bought from Lonza (Basel, Switzerland). Human recombinant IL2 (Proleukin ® ) was purchased from Novartis (Taipei, Taiwan, ROC). Zoledronic acid (ZometaTM) was purchased from Yungshin Pharm Ind. Co. Ltd. (Taipei, Taiwan, ROC). Alamar Blue™ Cell Viability Reagent, McCoy's 5a (for T24 cells), RPMI 1640 (for BFTC 905 cells), Lglutamate, fetal bovine serum and TrypLE Express Enzyme solution were bought from Gibco (ThermoFisher Scientific Inc., Waltham, MA, USA). Human monoclonal antibody isotype fluorescein isothiocyanate (FITC)-conjugated IgG1, peridinin-hlorophyll-protein (PerCP)-conjugated IgG1, allophycocyanin (APC)-conjugated IgG1, CD3-PerCP, brilliant violet (BV) 421-conjugated TCRγδ, APCconjugated natural killer group 2 member D (NKG2D), CD27-FITC, R-phyco-erythrin (PE)-cyanine 7 (Cy7)-conjugated CD45RA were all purchased from BD Biosciences (San Jose, CA, USA).
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