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Liberase

Liberase is a family of enzyme mixtures used for the isolation and purification of cells from various tissues, including pancreatic islets, liver, and adipose tissue.
These enzymes, derived from Clostridium histolyticum, are formulated to effectively digest the extracellular matrix, allowing for the gentle and efficient release of viable cells.
Liberase products are widely used in regenerative medicine research, cell transplantation, and tissue engineering applications.
Their ability to prseserve cell function and viability makes them a critical tool for advancing Liberase-related studies.

Most cited protocols related to «Liberase»

1
Optimized Isolation of Mouse Hepatocytes
Cited 59 times

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Publication 2020
Berries BLOOD Calcium Cannulation Cell Cycle Checkpoints Cells ChIP-Chip Collagenase Dental Anesthesia Digestion Extracellular Matrix Friend Hepatocyte Liberase Liver Mice, House Percoll Perfusion Veins, Portal Venae Cavae
2
Establishing Transgenic Melanoma Zebrafish
Cited 21 times
Transgenic melanoma zebrafish using the MiniCoopR system were created as
previously described (7 (link)). Briefly, a
plasmid was created in which the zebrafish mitfa promoter drives a zebrafish
MITF minigene devoid of introns. On the same plasmid was a second cassette in
which the mitfa promoter drives EGFP. Flanking both of these genes are Tol2
transposon arms. This plasmid was injected into fish with the following
genotype:
mitfa-BRAFV600E;p53−/−;mitfa−/−.
This strain of fish is devoid of all melanocytes (due to the
mitfa−/− mutation), but upon mosaic rescue with the
mitfa-MITF minigene will develop “patches” of rescued melanocytes,
some of which will go on to develop melanoma during adulthood. Because the
rescued melanocytes all contain the MiniCoopR plasmid, they will necessarily
also express mitfa-EGFP, resulting in melanomas which are entirely EGFP
positive. For the isolation of the cell lines, tumors were cleanly dissected
with a scalpel from melanoma bearing MiniCoopR fish and transferred to a small
petri dish containing 2 ml dissection medium (50% Ham's F12/50% DMEM, 10X
Pen/Strep, 0.075 mg/ml Liberase). They were then manually disaggregated for 30
minutes at room temperature. An inactivating solution (50% Ham's F12/50% DMEM,
10X Pen/Strep, 15% heat inactivated FCS) was then added, and the suspension
filtered 2-3X in a 40μM filter. This was then centrifuged for 5
minutes@500rcf, and resuspended in 500μl of complete zebrafish media (see
Supplemental
Methods for further details). This 500μl was then plated in a
single well of a 48-well plate that be been previously coated with
fibronectin.
Heilmann S., Ratnakumar K., Langdon E., Kansler E., Kim I., Campbell N.R., Perry E., McMahon A., Kaufman C., van Rooijen E., Lee W., Iacobuzio-Donahue C., Hynes R., Zon L., Xavier J, & White R. (2015). A quantitative system for studying metastasis using transparent zebrafish. Cancer research, 75(20), 4272-4282.
Publication 2015
Animals, Transgenic Arm, Upper Cell Separation Dissection Fishes Genes Hyperostosis, Diffuse Idiopathic Skeletal Introns Liberase Melanocyte Melanoma MITF protein, human Mutation Neoplasms Plasmids Strains Streptococcal Infections Zebrafish
3
Isolation and Characterization of Intestinal and Splenic Dendritic Cells
Cited 17 times
For LpDCs, total Lp cells after digestion with Liberase CI were passed through 70- and 40-μm cell strainers. Cells were resuspended in 1.077 g/cm3 iso-osmotic NycoPrep medium (Accurate Chemical & Scientific Corp.), and the low-density fraction was collected after centrifugation at 1,650 g for 15 min. Nycodenz gradient excludes debris and red blood cells and decreases lymphocyte numbers without changing the composition of the different subsets of LpDC (Fig. S5, available at http://www.jem.org/cgi/content/full/jem.20070663/DC1). Cells were washed and incubated with a mixture of mAb containing α−CD11c (HL-3), α−MHC II (AF6-120.1), α−CD16/32 (2.4G2), and α−CD103 mAb (2E7), as well as the non-DC components α−DX5 (DX5), α-NK1.1 (PK136), and α−B220 (RA3-6B2, all from eBioscience). DCs were defined as CD11c+MHCII+ cells and non-DCs were excluded when sorted by flow cytometry on a FACSVantage or FACSAria. In some experiments, CD103+ and CD103− DCs were separated. Purity was verified by flow cytometry on a FACSCalibur. I-Ab+CD11c+ cells were >90%.
For SpDCs, spleens were cut into fragments and digested by 100 mg/ml Liberase CI and 150 mg/ml DNase I, and then dissociated in Ca2+-free medium in the presence of EDTA. IAb+CD11c+ SpDCs were further purified for LpDCs, resulting in >98% purity. For phenotypic analysis, I-Ab+CD11c+ DCs were stained with fluorescent dye-conjugated α-CD11b (M1/70), α-CD103 (2E7; all from eBioscience), α-CD40 (3/23), α-CD80 (16-10A) (1 (link)), and α-CD86 (GL-1; all from BD Biosciences) mAb and analyzed by flow cytometry on a FACSCalibur or LSR II (BD Biosciences).
Sun C.M., Hall J.A., Blank R.B., Bouladoux N., Oukka M., Mora J.R, & Belkaid Y. (2007). Small intestine lamina propria dendritic cells promote de novo generation of Foxp3 T reg cells via retinoic acid. The Journal of Experimental Medicine, 204(8), 1775-1785.
Publication 2007
alpha HML-1 Cells Centrifugation Chief Cells, Gastric Deoxyribonucleases Edetic Acid Erythrocytes Flow Cytometry Fluorescent Dyes ITGAM protein, human Liberase Lymphocyte Count Nycodenz Osmosis Phenotype
4
Purification of Resting Macrophage Subsets
Cited 15 times
All cells were purified using the sorting protocol and antibodies listed on http://www.immgen.org. Cells were directly sorted from mouse tissues and were processed from tissue procurement to a second round of sorting into Trizol within 4 h using a Beckton-Dickinson Aria II instrument. Resting red pulp macrophages from the spleen were sorted after nonenzymatic disaggregation of the spleen and were identified as F4/80hi cells that lacked B220 and high levels of CD11c and MHC II34 (link),35 (link); macrophages from the resting peritoneum were collected in a peritoneal lavage and stained to identify CD115hi cells that were F4/80hi MHC II−; resting pulmonary macrophages were isolated from Liberase III-digested lungs (15 min. digest) and macrophages were identified as SiglecF+ CD11c+ cells with low levels of MHC II26 (link),27 (link); and resting brain microglial macrophages were sorted from Liberase III-digested, Percoll-gradient separated cells that were CD11b+ CD45lo F4/80lo11 (link). The Data Browser in the Immgen website is a resource for pdf files showing FACS dot plots that depict the purification strategies and purity after isolation of these and all other populations. A list of abbreviations used in the Immgen database relevant to macrophages and DCs can be found in Supplementary Note 1.
Gautier E.L., Shay T., Miller J., Greter M., Jakubzick C., Ivanov S., Helft J., Chow A., Elpek K.G., Gordonov S., Mazloom A.R., Ma’ayan A., Chua W.J., Hansen T.H., Turley S.J., Merad M, & Randolph G.J. (2012). Gene expression profiles and transcriptional regulatory pathways underlying mouse tissue macrophage identity and diversity. Nature immunology, 13(11), 1118-1128.
Publication 2012
Antibodies Brain Cells Dental Pulp isolation ITGAM protein, human Liberase Lung Macrophage Macrophages, Alveolar Macrophages, Peritoneal Microglia Mus NRG1 protein, human Percoll Peritoneal Lavage Population Group Spleen Tissue Procurement Tissues trizol
5
Isolation and Expansion of CD34+ Endothelial Progenitor Cells
Cited 14 times
Venous samples were minced and digested with Liberase 2 (Roche, Basel, Switzerland). For culture selection, 2×105 cells/mL were plated in the presence of Human Medium (HM; complete NeuroCult medium, StemCell Technologies Inc, Vancouver, British Columbia, Canada) that contained basic fibroblast growth factor (10 ng/mL) and epidermal growth factor (20 ng/mL; both from R&D Systems, Minneapolis, Minn) on uncoated wells (n=5).
CD34pos/CD31-negative (CD31neg) cells were isolated from saphenous vein digests either by magnetic bead–assisted cell sorting (MACS, Miltenyi Biotec, Bergisch Gladbach, Germany; n=6) or by fluorescence-activated cell sorting (n 2). Fluorescence-activated cell sorting was performed with a high-speed cell sorter (MoFlo, Beckman Coulter, Fullerton, Colo). Purity of the preparations was assessed by flow cytometry analysis. Differentiation/expansion was performed by seeding CD34posCD31neg cells obtained with each method on fibronectin-coated plates in endothelial growth medium (EGM2, Lonza, Basel, Switzerland) that contained 2% fetal bovine serum (Lonza).
Campagnolo P., Cesselli D., Zen A.A., Beltrami A.P., Kränkel N., Katare R., Angelini G., Emanueli C, & Madeddu P. (2010). Human Adult Vena Saphena Contains Perivascular Progenitor Cells Endowed With Clonogenic and Proangiogenic Potential. Circulation, 121(15), 1735-1745.
Publication 2010
Cells Endothelium Epidermal growth factor Epiphyseal Cartilage Fetal Bovine Serum Fibroblast Growth Factor 2 Fibronectins Flow Cytometry Homo sapiens Liberase Saphenous Vein Stem Cells Veins

Most recents protocols related to «Liberase»

1
Aortic Cell Suspension Preparation
To prepare aortic cell suspension, fresh descending aorta and aortic root fragments, harvested from Apoe−/− male C57BL/6 mice (18 weeks old) fed with a Western diet and normal diet, three in each group, were incubated by an enzyme mix with 0.2 mg/mL Liberase (Roche, 5,401,054,001) and 2 U/mL Elastase (Sigma-Aldrich, E1250), with HBSS as the solvent. Digestion was done by rotating at 37°C in an oven for an hour. The product was filtered through the 35 um strainer and washed with HBSS. Cells were collected by centrifugation at room temperature, 500 xg for 5 min. The supernatant was discarded and the cells resuspended with staining buffer (3% BSA and 1%NaN in PBS).
Each group of cell suspension mentioned above was divided into four parts for incubation with the following four antibodies.
Incubation was performed for 1 h at 4°C in darkness. Separate isotype controls for each antibody were also prepared. Secondary antibodies labeled with fluorescent dye were diluted with 3% BSA and used to resuspend cells at room temperature for 30 min in darkness.
Cells were washed with PBS by centrifugation at 400 g for 5 min twice. Finally, cells were resuspended with cold staining buffer (3% BSA and 1%NaN in PBS), the cell number was counted, and flow cytometry was performed. Cells were sorted by flow cytometry (CytoFLEX, Beckman Coulter) and analyzed with flow cytometer (CytoFLEX, Beckman Coulter) (version 2.0).
Wen J., Ling R., Chen R., Zhang S., Dai Y., Zhang T., Guo F., Wang Q., Wang G, & Jiang Y. (2023). Diversity of arterial cell and phenotypic heterogeneity induced by high-fat and high-cholesterol diet. Frontiers in Cell and Developmental Biology, 11, 971091.
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Publication 2023
Antibodies Aorta Aortic Root ApoE protein, human Buffers Cells Centrifugation Cold Temperature Darkness Descending Aorta Diet Digestion Enzymes Flow Cytometry Fluorescent Dyes Hemoglobin, Sickle Immunoglobulin Isotypes Immunoglobulins Liberase Males Mice, Inbred C57BL Pancreatic Elastase Solvents Strains
2
Isolation and Characterization of Peritoneal Cells
Tissues were separated, minced, and incubated in PBS containing 0.2 U/ml of Liberase TM (Roche) and 20 μg/ml DNase I (Roche) in the presence of calcium and magnesium. After digestion, the suspension was further mechanically disrupted by pipetting and filtered through 70-μm cell strainer. Peritoneal exudate cells were harvested by injecting 8 ml of PBS containing 10% fetal bovine serum and 2 mM EDTA into peritoneal cavity. Single-cell suspensions were preincubated with antibody against CD16/32 to block FcγRII/III receptors and stained on ice for 10 min with antibodies conjugated with fluorochrome. Flow cytometry was performed on an LSRFortessa (BD Biosciences), and aldehyde dehydrogenase activity was determined by ALDEFLUOR (StemCell Technologies).
Yoshihara T, & Okabe Y. (2023). Aldh1a2+ fibroblastic reticular cells regulate lymphocyte recruitment in omental milky spots. The Journal of Experimental Medicine, 220(5), e20221813.
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Publication 2023
Antibodies Ascites Calcium Cardiac Arrest CD32 Antigens Cells Dehydrogenase, Aldehyde Deoxyribonucleases Digestion Edetic Acid Fetal Bovine Serum Flow Cytometry Fluorescent Dyes Immunoglobulins Liberase Magnesium Peritoneal Cavity Stem Cells Tissues
3
Isolation of Lung, Adipose, and Liver Cells
Cells of the lung, adipose tissue and liver were isolated by enzymatic digestion as follows:
Lung: Lungs were perfused with PBS from the heart, isolated and minced first with scissors, followed by the gentleMACS program m_lung_01-02, digested 30 min at 37 °C on an orbital shaker with 0.15 WünschU/mg Liberase (Cat# 5401020001, Roche) and 0.1 mg/mL DNase I, and homogenized by using gentleMACS program m_lung_02_01. Lung cells were enriched for leukocytes by Percoll gradient (40% /70%, 600 g, 20 min, minimal brake). Gating see Additional file 1: Figure S2A.
Adipose tissue: Epididymal adipose tissue was minced and digested with 1.5 mg/mL collagenase IV (Cat# LS004189, Worthington), 10 mM HEPES and 8.25 µg/mL DNase I for 20–25 min on a thermomixer with 400 rpm. Erythrocytes were removed by red cell lysis buffer (154 mM NH4Cl, 10 mM KHCO3, 0.1 mM EDTA). Adipose tissue macrophages were gated as CD45+F4/80+CD11b+ and subdivided into subpopulations double negative (DN), pro-inflammatory M1a (CD11c+CD206−) and M1b (CD11c+CD206int), and anti-inflammatory M2 (CD11c− to lowCD206+). Gating see Additional file 1: Figure S2B.
Liver macrophages: One liver lobe was minced and digested using 1.5 mg/mL collagenase IV, 10 mM HEPES and 8.25 µg/mL DNase I, on a thermomixer with 400 rpm. After 15 min, the tissue was mechanistically dissected by pipetting up and down with a 1 mL pipette, and digested for another 15 min, followed by filtration and Percoll gradient. Gating see Additional file 1: Figure S2C.
Cell analysis was performed on a FACS LSRII Fortessa (BD Biosciences). Acquired data were analyzed using FlowJo software (Version 9.9 or higher), TreeStar Inc. Ashland, OR, USA).
From Biolegend, we obtained antibodies against CD11b (M1/70), CD11c (N418), MHCII (M5/114.14.2), Ly6C (HK1.4), CD45 (30-F11), F4/80 (BM8), CD103 (2E7), CD24 (M1/69), CD64 (X54-5/7.1), CD3 (145-2C11), CD19 (6D5), NK1.1 (PL136), Ly6G (1A8) and CD206 (C068C2). mAb for CCR2 (475,301) was purchased from R&D. mAb for Siglec F (E50-2440) was obtained from BD. For further details see Additional file 1: Table S3.
Bosch A.J., Rohm T.V., AlAsfoor S., Low A.J., Keller L., Baumann Z., Parayil N., Stawiski M., Rachid L., Dervos T., Mitrovic S., Meier D.T, & Cavelti-Weder C. (2023). Lung versus gut exposure to air pollution particles differentially affect metabolic health in mice. Particle and Fibre Toxicology, 20, 7.
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Publication 2023
alpha HML-1 Anti-Inflammatory Agents Antibodies Buffers Cells Collagenase Deoxyribonuclease I Deoxyribonucleases Digestion Edetic Acid Enzymes Epididymis Erythrocytes Filtration Heart HEPES Inflammation ITGAM protein, human Kupffer Cells Leukocytes Liberase Liver Lung Macrophage Mucolipidosis Type IV Mycoplasma pneumoniae Percoll Population Group potassium bicarbonate Sialic Acid Binding Immunoglobulin-like Lectins Tissue, Adipose Tissues
4
Characterization of Adipose Tissue and PBMC in Bariatric Patients
Subcutaneous adipose tissue and visceral adipose tissue biopsies obtained from bariatric surgery were immediately stored in Krebs-Ringer Phosphate (KRP) buffer until processing. The biopsies were cut into pieces and enzymatically digested with collagenase Type I (Life Technologies) for 1 hour at 37°C with constant shaking. The subcutaneous adipose tissue from liposuction was washed in 0.9% NaCl, diluted in KRP buffer and digested with Liberase (Roche). The dissolved tissues were filtered, and the stromal vascular cells (SVC) were isolated from the mature adipocytes and washed with Phosphate Buffered Saline (PBS). The liposuction SVC was freshly stained, whereas the SVC from biopsies were preserved in freezing media containing FBS and 10% DMSO and stored in liquid nitrogen until further flow cytometry experiments. Peripheral blood mononuclear cells (PBMC) were isolated from blood of bariatric surgery patients and healthy donors using density gradient centrifugation, as previously described (33 (link)).
Haugstøyl M.E., Cornillet M., Strand K., Stiglund N., Sun D., Lawrence-Archer L., Hjellestad I.D., Busch C., Mellgren G., Björkström N.K, & Fernø J. (2023). Phenotypic diversity of human adipose tissue-resident NK cells in obesity. Frontiers in Immunology, 14, 1130370.
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Publication 2023
Adipocytes Bariatric Surgery Biopsy BLOOD Blood Vessel Centrifugation, Density Gradient Collagenase, Clostridium histolyticum Donors Flow Cytometry Liberase Nitrogen Normal Saline Patients PBMC Peripheral Blood Mononuclear Cells Phosphates Saline Solution Stromal Cells Subcutaneous Fat Suction Lipectomy Sulfoxide, Dimethyl Tissues Visceral Fat
5
Isolation of Cadaveric Islet Cells
hPIs used in this study were isolated from six cadaveric multiorgan donors, in particular three women and three men, donor age 52.6 ± 3.6 years, BMI 25.5 ± 2.6 kg/m2, islets purity 82.5 ± 5.2%, according to the procedure previously described (Ricordi et al., 1988 (link); Petrelli et al., 2011 (link)). The overall protocol has been approved by the Niguarda Cà Granda Ethics Board. Islets were isolated using the automated method previously described (Ricordi et al., 1988 (link)). Pancreata were obtained from multiorgan cadaveric donors utilizing cold perfusion. Exclusion and inclusion criteria were applied based on the Italian Guidelines. Briefly, pancreata were digested by a cold enzymatic blend solution of collagenase and thermolysin (Liberase MTF GMP Grade kit, Roche Diagnostics, Mannheim, Germany) reconstituted in Hank’s Balanced Salt Solution (HBSS, Euroclone, Italy) with 25 mM of HEPES. Subsequently, islets were purified with discontinuous polysucrose solutions at decreasing density 1.132, 1.108, 1.096, 1.060 and 1.037 g/L (Mediatech-Cellgro, VA, United States). Islets were counted by dithizone staining islet equivalent (IEQ) method (see “Dithizone staining” section) and they were cultured at 24°C, 20% O2, 5% CO2 in a humidified atmosphere in MIAMI Medium #1A (Mediatech-Cellgro, VA, USA) supplemented with Ciprofloxacin (Fresenius Kabi, Verona, Italy), or in serum-free medium in the presence of basic fibroblast growth factor (bFGF, PeproTech) and epidermal growth factor (EGF, PeproTech) at final concentrations of 10 ng/ml and 20 ng/ml, respectively.
Marchini A., Ciulla M.G., Antonioli B., Agnoli A., Bovio U., Visnoviz V., Bertuzzi F, & Gelain F. (2023). Long-term cultures of human pancreatic islets in self-assembling peptides hydrogels. Frontiers in Bioengineering and Biotechnology, 11, 1105157.
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Publication 2023
Atmosphere Ciprofloxacin Cold Temperature Collagenase Diagnosis Dithizone Donors Enzymes Epidermal growth factor Fibroblast Growth Factor 2 Hemoglobin, Sickle HEPES Liberase Pancreas Perfusion Serum Sodium Chloride Thermolysin Tissue Donors Woman

Top products related to «Liberase»

1
Liberase by Roche
Sourced in Switzerland, Germany, United States, United Kingdom, France, Spain, Sweden, Canada, Norway, Austria, Japan, China, Denmark
Liberase is a laboratory enzyme product manufactured by Roche. It is designed to facilitate the isolation and dissociation of cells and tissues for various research and diagnostic applications.
2
Liberase tl by Roche
Sourced in Switzerland, United States, Germany, United Kingdom
Liberase TL is a digestive enzyme mixture used in laboratory applications. It is designed to dissociate tissue samples into single cells or small cell clusters. The product consists of a blend of purified enzymes that effectively break down the extracellular matrix, allowing for the isolation of viable cells from various tissues.
3
Dnase 1 by Roche
Sourced in United States, Switzerland, Germany, Japan, United Kingdom, France, Canada, Italy, Macao, China, Australia, Belgium, Israel, Sweden, Spain, Austria
DNase I is a lab equipment product that serves as an enzyme used for cleaving DNA molecules. It functions by catalyzing the hydrolytic cleavage of phosphodiester bonds in the DNA backbone, effectively breaking down DNA strands.
4
Dnase 1 by Merck Group
Sourced in United States, Germany, Switzerland, United Kingdom, Italy, Japan, Macao, Canada, Sao Tome and Principe, China, France, Australia, Spain, Belgium, Netherlands, Israel, Sweden, India
DNase I is a laboratory enzyme that functions to degrade DNA molecules. It catalyzes the hydrolytic cleavage of phosphodiester linkages in the DNA backbone, effectively breaking down DNA strands.
5
Liberase by Merck Group
Sourced in United States, Germany
Liberase is a proprietary enzyme blend designed for the gentle and effective dissociation of tissues and cells. It is a versatile tool used in a variety of research and clinical applications, such as cell isolation, tissue engineering, and regenerative medicine.
6
Dnase by Roche
Sourced in United States, Switzerland, Germany, United Kingdom, Japan, France, Australia, Canada, Sweden, Belgium
DNase is a type of enzyme that catalyzes the degradation of DNA. It is used in various laboratory techniques to remove unwanted DNA from samples.
7
Dnase by Merck Group
Sourced in United States, Germany, United Kingdom, Switzerland, France, Italy, Canada, Spain, China, Sao Tome and Principe, Jersey, Australia, Macao
DNase is a type of enzyme that catalyzes the degradation of DNA. It functions by cleaving the phosphodiester bonds in the DNA backbone, effectively breaking down DNA molecules into smaller fragments.
8
Liberase tl by Merck Group
Sourced in United States, Germany, France
Liberase TL is a lab equipment product manufactured by Merck Group. It is an enzyme blend designed for effective tissue dissociation and cell isolation. The core function of Liberase TL is to facilitate the extraction and separation of cells from various tissue types, enabling further research and analysis.
9
Liberase th by Roche
Sourced in Germany, United States, Switzerland
Liberase TH is a digestive enzyme reagent used in cell isolation and tissue dissociation applications. It contains a proprietary blend of collagenase enzymes that effectively break down the extracellular matrix, enabling the release of cells from a variety of tissues.

More about "Liberase"

Discover the versatility of Liberase, a family of enzyme mixtures that have revolutionized cell isolation and purification processes in regenerative medicine, cell transplantation, and tissue engineering.
Derived from the bacterium Clostridium histolyticum, these enzymes effectively digest the extracellular matrix, allowing for the gentle and efficient release of viable cells from various tissues, including pancreatic islets, liver, and adipose tissue.
Liberase products, such as Liberase TL and Liberase TH, are widely used in research due to their ability to preserve cell function and viability, making them a critical tool for advancing Liberase-related studies.
These enzyme mixtures, which may also incorporate DNase I or DNase, are formulated to optimize the isolation and purification of cells, ensuring reproducibility and accuracy in your research.
Explore the power of AI-driven protocol comparisons with PubCompare.ai, which can help you quickly locate the best Liberase-related protocols from literature, pre-prints, and patents.
This intelligent platform can enhance your Liberase research by assisting you in making informed decisions and elevating your work to new heights.
Experienence the power of AI-assisted optimization and take your Liberase studies to the next level.