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cryostor cs10 freeze medium  (BioLife Solutions)

 
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    BioLife Solutions cryostor cs10 freeze medium
    High post-thawing viability and proliferative capacity of G-rex expanded HSPC-NK cells cryopreserved using an optimized freezing and thawing protocol A Overview of the GMP-compliant generation of NK cells from HSPCs in a G-rex scalable bioreactor using six standardized medium additions. B Fold expansion at day 35. C Absolute number of CD34 + HSPCs at day 0 and CD56 + HSPC-NK cells at day 35. D Composition of the HSPC-NK cell product. E Trypan blue-based post-thawing viability of HSPC-NK cells frozen in cryobags. F Recovery of viable cells post-thawing compared to pre-freezing dosage of bag-frozen cells. G HSPC-NK cell dosage post-thawing. H Stability of thawed HSPC-NK cells in <t>CryoStor</t> <t>CS10</t> for a 30-min timespan. I Post-thawing viability following twelve months of storage in the liquid nitrogen vapor phase, assessed using cryovials. Stability study ongoing till five years post-freezing. J Absolute number of viable fresh or cryopreserved HSPC-NK cells cultured up to seven days in 50 ng/ml IL-15. K Proliferation score, defined as the fold change reduction in MFI of Proliferation Dye eFluor450 compared to day zero, of fresh and cryopreserved HSPC-NK cells following seven days of culture in 50 ng/ml IL-15. Donor 4 (▼) involves a non-GMP compliant pre-validation run. For donor 3 (∎), t = 0 represents t = 15 ( E - G ) and t = 30 represents t = 45 ( G ) due to technical issues. Bars in B, D-G, K represent the mean. Significance was evaluated using the Wilcoxon test for H, Friedman test with Dunn’s multiple comparison correction for I, and a two-way ANOVA with Šídák multiple comparison correction for K; no statistical significances were found
    Cryostor Cs10 Freeze Medium, supplied by BioLife Solutions, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cryostor+cs10+freeze+medium/cryostor+cs10/pmc12104124-74-12-17
    Average 90 stars, based on 1 article reviews
    cryostor cs10 freeze medium - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "Good manufacturing practice production of an off-the-shelf CD34 + progenitor-derived NK cell product with preserved anti-tumor functionality post-infusion in NOD/SCID/IL2Rg null mice"

    Article Title: Good manufacturing practice production of an off-the-shelf CD34 + progenitor-derived NK cell product with preserved anti-tumor functionality post-infusion in NOD/SCID/IL2Rg null mice

    Journal: Cellular and Molecular Life Sciences: CMLS

    doi: 10.1007/s00018-025-05727-4

    High post-thawing viability and proliferative capacity of G-rex expanded HSPC-NK cells cryopreserved using an optimized freezing and thawing protocol A Overview of the GMP-compliant generation of NK cells from HSPCs in a G-rex scalable bioreactor using six standardized medium additions. B Fold expansion at day 35. C Absolute number of CD34 + HSPCs at day 0 and CD56 + HSPC-NK cells at day 35. D Composition of the HSPC-NK cell product. E Trypan blue-based post-thawing viability of HSPC-NK cells frozen in cryobags. F Recovery of viable cells post-thawing compared to pre-freezing dosage of bag-frozen cells. G HSPC-NK cell dosage post-thawing. H Stability of thawed HSPC-NK cells in CryoStor CS10 for a 30-min timespan. I Post-thawing viability following twelve months of storage in the liquid nitrogen vapor phase, assessed using cryovials. Stability study ongoing till five years post-freezing. J Absolute number of viable fresh or cryopreserved HSPC-NK cells cultured up to seven days in 50 ng/ml IL-15. K Proliferation score, defined as the fold change reduction in MFI of Proliferation Dye eFluor450 compared to day zero, of fresh and cryopreserved HSPC-NK cells following seven days of culture in 50 ng/ml IL-15. Donor 4 (▼) involves a non-GMP compliant pre-validation run. For donor 3 (∎), t = 0 represents t = 15 ( E - G ) and t = 30 represents t = 45 ( G ) due to technical issues. Bars in B, D-G, K represent the mean. Significance was evaluated using the Wilcoxon test for H, Friedman test with Dunn’s multiple comparison correction for I, and a two-way ANOVA with Šídák multiple comparison correction for K; no statistical significances were found
    Figure Legend Snippet: High post-thawing viability and proliferative capacity of G-rex expanded HSPC-NK cells cryopreserved using an optimized freezing and thawing protocol A Overview of the GMP-compliant generation of NK cells from HSPCs in a G-rex scalable bioreactor using six standardized medium additions. B Fold expansion at day 35. C Absolute number of CD34 + HSPCs at day 0 and CD56 + HSPC-NK cells at day 35. D Composition of the HSPC-NK cell product. E Trypan blue-based post-thawing viability of HSPC-NK cells frozen in cryobags. F Recovery of viable cells post-thawing compared to pre-freezing dosage of bag-frozen cells. G HSPC-NK cell dosage post-thawing. H Stability of thawed HSPC-NK cells in CryoStor CS10 for a 30-min timespan. I Post-thawing viability following twelve months of storage in the liquid nitrogen vapor phase, assessed using cryovials. Stability study ongoing till five years post-freezing. J Absolute number of viable fresh or cryopreserved HSPC-NK cells cultured up to seven days in 50 ng/ml IL-15. K Proliferation score, defined as the fold change reduction in MFI of Proliferation Dye eFluor450 compared to day zero, of fresh and cryopreserved HSPC-NK cells following seven days of culture in 50 ng/ml IL-15. Donor 4 (▼) involves a non-GMP compliant pre-validation run. For donor 3 (∎), t = 0 represents t = 15 ( E - G ) and t = 30 represents t = 45 ( G ) due to technical issues. Bars in B, D-G, K represent the mean. Significance was evaluated using the Wilcoxon test for H, Friedman test with Dunn’s multiple comparison correction for I, and a two-way ANOVA with Šídák multiple comparison correction for K; no statistical significances were found

    Techniques Used: Cell Culture, Biomarker Discovery, Comparison

    Related Articles

    Concentration Assay:

    Article Title: Good manufacturing practice production of an off-the-shelf CD34 + progenitor-derived NK cell product with preserved anti-tumor functionality post-infusion in NOD/SCID/IL2Rg null mice
    Article Snippet: .. Afterwards, cells were cryopreserved in either room temperature (RT) or 4 °C pre-cooled CryoStor CS10 freeze medium (BioLife Solutions, 210102) at a concentration of 10-100 × 10 6 cells/ml in Cryotube vials (cryovials; ThermoFisher Scientific, 363401 or Greiner Bio-One, 126263) or CryoMACS freezing bags 50 (Miltenyi Biotech, 200-074-400). ..

    Article Title: Good manufacturing practice production of an off-the-shelf CD34 + progenitor-derived NK cell product with preserved anti-tumor functionality post-infusion in NOD/SCID/IL2Rg null mice.
    Article Snippet: .. Afterwards, cells were cryopreserved in either room temperature (RT) or 4 °C pre-cooled CryoStor CS10 freeze medium (BioLife Solutions, 210102) at a concentration of 10-100 × 106 cells/ml in Cryotube vials (cryovials; ThermoFisher Scientific, 363401 or Greiner Bio-One, 126263) or CryoMACS freezing bags 50 (Miltenyi Biotech, 200-074-400). ..

    Isolation:

    Article Title: Ex vivo -expanded highly pure ABCB5 + mesenchymal stromal cells as Good Manufacturing Practice-compliant autologous advanced therapy medicinal product for clinical use: process validation and first in-human data
    Article Snippet: After seven passages, ABCB5 + MSCs were isolated using magnetic beads (micromer TC1 epoxy; Micromod, Rostock, Germany) coated with a mouse anti-human monoclonal antibody directed against sequence 493508 (RFGAYLIQAGRMTPEG) of ABCB5 extracellular loop three [ ] (antibody bulk production: Maine Biotechnology Services, Portland, ME, USA; antibody GMP purification: Bibitec, Bielefeld, Germany; virus depletion and safety evaluation of the antibody according to [ ]: Charles River, Erkrath, Germany). .. After enzymatic (TrypZean) detachment of the beads from the cell surface, the isolated ABCB5 + MSCs were cryopreserved in CryoStor CS10 freeze medium (BioLife Solutions, Bothell, WA, USA) and stored in the vapor phase of liquid nitrogen. ..

    Saline:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Recombinant:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Sterility:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Pore Size:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Transferring:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Cell Culture:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Microscopy:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..

    Blocking Assay:

    Article Title: Protocol to engineer apical-out airway organoids using suspension culture of human airway basal stem cell aggregates.
    Article Snippet: .. 1 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Dimethyl sulfoxide (DMSO) Sigma-Aldrich Cat# D4540 Fetal bovine serum Fisher Scientific Cat# FB12999102 Penicillin-Streptomycin Thermo Fisher Scientific Cat# 15140148 Bronchial epithelial cell growth basal medium Lonza Cat# CC-3171 BEGM Bronchial Epithelial SingleQuots Kit Lonza Cat# CC-4175 A 83-01 (inhibitor of TGF-b type 1 receptor kinase) Sigma-Aldrich Cat# SML0788 Y-27632 Dihydrochloride (inhibitor of Rhoassociated protein kinase) Cayman Chemical Cat# 129830-38-2 CHIR99021 (GSK-3b inhibitor), 10 mM Reprocell Cat# 04000402 DMH-1 (inhibitor of BMP4/SMAD signaling) Tocris Bioscience Cat# 4126 PneumaCult-ALI Medium Stemcell Technologies Cat# 05001 Heparin solution Stemcell Technologies Cat# 07980 Hydrocortisone stock solution Stemcell Technologies Cat# 07925 0.25% Trypsin-EDTA Gibco Cat# 25200056 Dulbecco’s phosphate-buffered saline (DPBS), 13 without calcium and magnesium Corning Cat# 21-031-CV CryoStor CS10 Freeze Medium Biolife Solutions Cat# 210102 Corning CoolCell Container Corning Cat# 432000 Recombinant human interleukin-13 (IL-13) Peprotech Cat# 200-13 Growth factor reduced Matrigel Corning Cat# CB 40230 Triton X-100 Sigma-Aldrich Cat# X100-500mL 4% Paraformaldehyde Santa Cruz Biotechnology Cat# sc-281692 Tween-20 Sigma-Aldrich Cat# P2287 Bovine serum albumin Sigma-Aldrich Cat# 9048-46-8 DAPI (40,6-diamidino-2-phenylindole) in H2O, 10 mg/mL Biotium Cat# 40043 Experimental models: Cell lines 804G rat bladder epithelial cells Gift from Dr. Hongmei Mou2 N/A Normal human bronchial epithelial cells without retinoic acid Lonza Cat# CC-2541 Other Nalgene rapid-flow sterile disposable filter units, 0.45-mm pore size Thermo Fisher Scientific Cat# 124-0045 Wide-orifice pipette tips, 200 mL Axygen T-205-WB-C-R-S 75-cm2 cell culture flask Greiner Bio-One Cat# 658175 175-cm2 cell culture flask Greiner Bio-One Cat# 660175 96-well cell-repellent, U-bottom microplate Greiner Bio-One Cat# 650970 96-well cell-culture treated, flat-bottom microplate Greiner Bio-One Cat# 655180 Circular cover glass (25 mm) Fisher Scientific Cat# 12-545-102 Attofluor cell chamber, for microscopy Thermo Fisher Scientific Cat# A7816 Water bath Fisher Scientific Cat# FSGPD10 Incubator for cell culture Fisher Scientific Cat# 11676604 LSM 700 Confocal Microscope Zeiss N/A Sorvall Legend X1 Centrifuge Thermo Fisher Scientific Cat# 75004220 M-20 Microplate Swinging Bucket Rotor Thermo Fisher Scientific Cat# 75003624 Heat Block VWR International Cat#13259-032 ECLIPSE Ts2 Inverted Cell Culture Microscope Nikon N/A ll OPEN ACCESS Protocol ..



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    High post-thawing viability and proliferative capacity of G-rex expanded HSPC-NK cells cryopreserved using an optimized freezing and thawing protocol A Overview of the GMP-compliant generation of NK cells from HSPCs in a G-rex scalable bioreactor using six standardized medium additions. B Fold expansion at day 35. C Absolute number of CD34 + HSPCs at day 0 and CD56 + HSPC-NK cells at day 35. D Composition of the HSPC-NK cell product. E Trypan blue-based post-thawing viability of HSPC-NK cells frozen in cryobags. F Recovery of viable cells post-thawing compared to pre-freezing dosage of bag-frozen cells. G HSPC-NK cell dosage post-thawing. H Stability of thawed HSPC-NK cells in CryoStor CS10 for a 30-min timespan. I Post-thawing viability following twelve months of storage in the liquid nitrogen vapor phase, assessed using cryovials. Stability study ongoing till five years post-freezing. J Absolute number of viable fresh or cryopreserved HSPC-NK cells cultured up to seven days in 50 ng/ml IL-15. K Proliferation score, defined as the fold change reduction in MFI of Proliferation Dye eFluor450 compared to day zero, of fresh and cryopreserved HSPC-NK cells following seven days of culture in 50 ng/ml IL-15. Donor 4 (▼) involves a non-GMP compliant pre-validation run. For donor 3 (∎), t = 0 represents t = 15 ( E - G ) and t = 30 represents t = 45 ( G ) due to technical issues. Bars in B, D-G, K represent the mean. Significance was evaluated using the Wilcoxon test for H, Friedman test with Dunn’s multiple comparison correction for I, and a two-way ANOVA with Šídák multiple comparison correction for K; no statistical significances were found

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: Good manufacturing practice production of an off-the-shelf CD34 + progenitor-derived NK cell product with preserved anti-tumor functionality post-infusion in NOD/SCID/IL2Rg null mice

    doi: 10.1007/s00018-025-05727-4

    Figure Lengend Snippet: High post-thawing viability and proliferative capacity of G-rex expanded HSPC-NK cells cryopreserved using an optimized freezing and thawing protocol A Overview of the GMP-compliant generation of NK cells from HSPCs in a G-rex scalable bioreactor using six standardized medium additions. B Fold expansion at day 35. C Absolute number of CD34 + HSPCs at day 0 and CD56 + HSPC-NK cells at day 35. D Composition of the HSPC-NK cell product. E Trypan blue-based post-thawing viability of HSPC-NK cells frozen in cryobags. F Recovery of viable cells post-thawing compared to pre-freezing dosage of bag-frozen cells. G HSPC-NK cell dosage post-thawing. H Stability of thawed HSPC-NK cells in CryoStor CS10 for a 30-min timespan. I Post-thawing viability following twelve months of storage in the liquid nitrogen vapor phase, assessed using cryovials. Stability study ongoing till five years post-freezing. J Absolute number of viable fresh or cryopreserved HSPC-NK cells cultured up to seven days in 50 ng/ml IL-15. K Proliferation score, defined as the fold change reduction in MFI of Proliferation Dye eFluor450 compared to day zero, of fresh and cryopreserved HSPC-NK cells following seven days of culture in 50 ng/ml IL-15. Donor 4 (▼) involves a non-GMP compliant pre-validation run. For donor 3 (∎), t = 0 represents t = 15 ( E - G ) and t = 30 represents t = 45 ( G ) due to technical issues. Bars in B, D-G, K represent the mean. Significance was evaluated using the Wilcoxon test for H, Friedman test with Dunn’s multiple comparison correction for I, and a two-way ANOVA with Šídák multiple comparison correction for K; no statistical significances were found

    Article Snippet: Afterwards, cells were cryopreserved in either room temperature (RT) or 4 °C pre-cooled CryoStor CS10 freeze medium (BioLife Solutions, 210102) at a concentration of 10-100 × 10 6 cells/ml in Cryotube vials (cryovials; ThermoFisher Scientific, 363401 or Greiner Bio-One, 126263) or CryoMACS freezing bags 50 (Miltenyi Biotech, 200-074-400).

    Techniques: Cell Culture, Biomarker Discovery, Comparison