Log In Sign up
The largest database of trusted experimental protocols
> Chemicals & Drugs > Amino Acid > Dispase

Dispase

Dispase is a proteolytic enzyme used in cell and tissue dissociation procedures.
It is derived from Bacillus polymyxa and has the ability to cleave a variety of extracellular matrix components, making it useful for isolating cells from tissues.
Dispase has been widely employed in research involving stem cells, tissue engineering, and regenerative medicine.
When compared to other dissociation enzymes, Dispase often provides a gentler alternative that can help maintain cell viability and functionality.
Researchers utilzing Dispase should be aware of the potential for variable activity and effectiveness, depending on factors such as source, lot, and experimental conditions.
Optimizing Dispase protocols through careful comparison can enhance reproducibility and unlock new insights in Dispase-based research.

Most cited protocols related to «Dispase»

1
Cerebral Organoid Generation from Human Pluripotent Stem Cells
Cited 74 times
Human H9 ES (WA09) were obtained from WiCell at passage 26 with verified normal karyotype and contamination-free. iPS cells were obtained from System Biosciences (SC101A-1) verified pluripotent and contamination free. All human PSC lines were regularly checked and confirmed negative for mycoplasma. PSCs were maintained on CF-1 gamma irradiated MEFs (Global Stem) according to WiCell protocols. On day 0 of organoid culture, ESCs or iPSCs less than passage 50 were dissociated from MEFs by dispase treatment and MEFs were removed by gravity separation of stem cell colonies from MEFs before trypsinization of stem cells to generate single cells. 4500 cells were then plated in each well of an ultra-low binding 96-well plate (Corning) in hES media with low bFGF (5-fold reduced) and 50uM ROCK inhibitor49 (link) (Calbiochem).
EBs were fed every other day for 6 days then transferred to low adhesion 24-well plates (Corning) in neural induction media containing DMEM/F12, 1:100 N2 supplement (Invitrogen), Glutamax (Invitrogen), MEM-NEAA, and 1ug/ml Heparin50 (link) (Sigma). These began forming neuroepithelial tissues, which were fed every other day for 5 days. On Day 11 of the protocol, tissues were transferred to droplets of Matrigel (BD Biosciences) by pipetting into cold Matrigel on a sheet of Parafilm with small 3mm dimples. These droplets were allowed to gel at 37C and were subsequently removed from the Parafilm and grown in differentiation media containing a 1:1 mixture of DMEM/F12 and Neurobasal containing 1:200 N2 supplement (Invitrogen), 1:100 B27 supplement without vitamin A (Invitrogen), 3.5ul/L 2-mercaptoethanol, 1:4000 insulin (Sigma), 1:100 Glutamax (Invitrogen), 1:200 MEM-NEAA.
After 4 days of stationary growth, the tissue droplets were transferred to a spinning bioreactor containing differentiation media as above except B27 supplement with vitamin A (Invitrogen) was used. Since retinoic acid has been shown to be important for neuronal differentiation in vivo52 (link), we included it in the final media used to differentiate the cerebral organoids.
Lancaster M.A., Renner M., Martin C.A., Wenzel D., Bicknell L.S., Hurles M.E., Homfray T., Penninger J.M., Jackson A.P, & Knoblich J.A. (2013). Cerebral organoids model human brain development and microcephaly. Nature, 501(7467), 10.1038/nature12517.
Publication 2013
2-Mercaptoethanol Bioreactors Cells Cell Separation Common Cold Dietary Supplements dispase Enhanced S-Cone Syndrome Gamma Rays Gravity Homo sapiens Induced Pluripotent Stem Cells Insulin Karyotyping matrigel Mycoplasma Nervousness Neurons Organoids Pancreatic Stellate Cells Stem, Plant Stem Cells Tissues Tretinoin Vitamin A
2
Isolation of Normal and Neoplastic Pancreatic Ducts
Cited 72 times

Protocol full text hidden due to copyright restrictions

Open the protocol to access the free full text link

Publication 2014
Collagenase Dietary Fiber Digestion dispase Enzymes Growth Factor matrigel Neoplasm Metastasis Neoplasms Pancreas Pancreatic Duct RNA-Seq Tissues
3
Neural Induction and Differentiation of hESCs/iPSCs
Cited 66 times
hESCs or iPSCs were isolated from MEFs following dissociation to single cells with Accutase (Innovative Cell Technologies) by a 1 hr pre-plate on gelatin-coated dishes in hESC medium supplemented with 10 ng/ml FGF2 and 10 μM ROCK inhibitor (Calbiochem). The non-adherent pluripotent stem cells were harvested and plated on Matrigel (BD) coated 12-well plates in MEF-conditioned hESC medium with 10 ng/ml FGF2. Once the cell culture reached 95% confluence, neural induction was initiated by changing the culture medium to a culture medium that supports neural induction, neurogenesis and neuronal differentiation (referred to as 3N medium), a 1:1 mixture of N2- and B27-containing media. N2 medium: DMEM/F12, N2 (GIBCO), 5 μg/ml Insulin, 1mM L-Glutamine, 100 μm non-essential amino acids, 100 μM 2-mercaptoethanol, 50 U/ml Penicillin and 50 mg/ml Streptomycin; B27 medium: Neurobasal (Invitrogen), B27 with or without vitamin A (GIBCO), 200 mM Glutamine, 50 U/ml Penicillin and 50 mg/ml Streptomycin. 3N medium was supplemented with either 1 μm Dorsomorphin (Tocris) or 500 ng/ml mouse Noggin-CF chimera (R&D Systems), and 10 μm SB431542 (Tocris) to inhibit TGFβ signaling during neural induction 19 (link). Cells were maintained in this medium for 8-11 days, during which time the efficiency of neural induction was monitored by the appearance of cells with characteristic neuroepithelial cell morphology. Neuroepithelial cells were harvested by dissociation with Dispase and replated in 3N medium including 20 ng/ml FGF2 on poly-ornithine and laminin-coated plastic plates. After a further 2 days, FGF2 was withdrawn to promote differentiation. Cultures were passaged once more with Accutase, replated at 50,000 cells/cm2 on poly-ornithine and laminin-coated plastic plates in 3N medium and maintained for up to 100 days with a medium change every other day.
For quantitative RT-PCR, total RNA was isolated from three cultures at each timepoint (days 5, 10, 15, 20 and 25) (Trizol, Sigma). Total RNA was reverse-transcribed and used for quantitative RT-PCR with primers specific to Foxg1 and Tbr2 using the Applied Biosystems 7000 system. Semi-quantitative RT-PCR with primers for Emx1, Dlx1, Nkx2.1, HoxB4 and Isl1 was carried out according to standard techniques on first strand, random-primed cDNA generated from total RNA extracted from cultures grown in the presence or absence of purmorphamine.
Shi Y., Kirwan P., Smith J., Robinson H.P, & Livesey F.J. (2012). Human cerebral cortex development from pluripotent stem cells to functional excitatory synapses. Nature neuroscience, 15(3), 10.1038/nn.3041.
Publication 2012
2-Mercaptoethanol 4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide accutase Amino Acids, Essential Cardiac Arrest Cell Culture Techniques Cells Chimera Culture Media, Conditioned dispase DNA, Complementary dorsomorphin Fibroblast Growth Factor 2 Gelatins Glutamine Human Embryonic Stem Cells Hyperostosis, Diffuse Idiopathic Skeletal Induced Pluripotent Stem Cells Insulin Laminin matrigel Mus Nervousness Neuroepithelial Cells Neurogenesis Neurons NKX2-1 protein, human noggin protein Oligonucleotide Primers Ornithine Penicillins Pluripotent Stem Cells Poly A purmorphamine Reverse Transcriptase Polymerase Chain Reaction Streptomycin Transforming Growth Factor beta trizol Vitamin A
4
Neural Induction of Pluripotent Stem Cells
Cited 65 times
For dissociating intact colonies of pluripotent stem cells from the layer of DR4 feeders, hiPSCs were exposed to a low concentration of dispase (Invitrogen: 17105-041; 0.7 mg/ml) for ~30 min. Suspended colonies were subsequently transferred into ultra-low-attachment 100 mm plastic plates (Corning) in hiPSC medium without FGF2. For the first 24 h (day 0), the medium was supplemented with the ROCK inhibitor Y-27632 (EMD Chemicals). For neural induction, dorsomorphin (also known as compound C; Sigma 10 μM) and SB-431542 (Tocris, 10 μM) were added to the medium for the first five days. On the sixth day in suspension, the floating spheroids were moved to neural medium (NM) containing Neurobasal (Invitrogen: 10888), B-27 serum substitute without vitamin A (Invitrogen: 12587), GlutaMax (Invitrogen, 1:100), 100 U/ml penicillin and 100 μl streptomycin (Invitrogen). The NM was supplemented with 20 ng/ml FGF2 (R&D Systems) and 20 ng/ml EGF (R&D Systems) for 19 days with daily medium change in the first 10 days, and every other day for the subsequent 9 days. To promote differentiation of the neural progenitors into neurons, FGF2 and EGF were replaced with 20 ng/ml BDNF (Peprotech) and 20 ng/ml NT3 (Peprotech) starting at day 25, while from day 43 onwards only NM without growth factors was used for medium changes every four days.
Paşca A.M., Sloan S.A., Clarke L.E., Tian Y., Makinson C.D., Huber N., Kim C.H., Park J.Y., O’Rourke N.A., Nguyen K.D., Smith S.J., Huguenard J.R., Geschwind D.H., Barres B.A, & Paşca S.P. (2015). Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture. Nature methods, 12(7), 671-678.
Publication 2015
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide dispase dorsomorphin Feeder Cell Layers Fibroblast Growth Factor 2 Growth Factor Human Induced Pluripotent Stem Cells Nervousness Neurons Penicillins Pluripotent Stem Cells Serum Streptomycin Vitamin A Y 27632
5
Isolation of Murine Skeletal Muscle Progenitors
Cited 41 times
Tibialis anterior muscles of mice were subjected to enzymatic dissociation (first collagenase 0.2% and then dispase (0.04 units ml−1), Sigma) for 90 min, after which non-muscle tissue was gently removed under a dissection microscope. The cell suspension was filtered through a 70 µm nylon filter (Falcon) and incubated with the following biotinylated antibodies: CD45, CD11b, CD31 and Sca1 (BD Bioscience). Streptavidin beads (Miltenyi Biotech) were then added to the cells together with the following antibodies: integrin-α7–phycoerythrin (PE; a gift from F. Rossi) and CD34–Alexa647 (eBioscience), after which magnetic depletion of biotin-positive cells was performed. The (CD45−CD11b−CD31−Sca1−)CD34+integrin-α7+ population was then fractionated twice by flow cytometry (DIVA-Van, Becton-Dickinson). Primary myoblasts were isolated as described previously20 (link).
Sacco A., Doyonnas R., Kraft P., Vitorovic S, & Blau H.M. (2008). Self-renewal and expansion of single transplanted muscle stem cells. Nature, 456(7221), 502-506.
Publication 2008
Alexa Fluor 647 Antibodies Biotin CASP3 protein, human Cells dispase Dissection Enzymes Flow Cytometry Integrins ITGAM protein, human Microscopy Mus Muscle Tissue Myoblasts Neutrophil Collagenase Nylons Phycoerythrin Streptavidin Tibial Muscle, Anterior

Most recents protocols related to «Dispase»

1
Mesodermal ECM Disruption Protocol
Not available on PMC !
After removing the ectoderm or endoderm mechanically, 10µl of ECM degrading enzyme Dispase-ii (Roche) was applied to the exposed mesodermal surface for 5 minutes, washed three times using 0.5ml 1XPBS, before preparing the 200mn slices. In parallel, embryo slices were fixed and examined by immunofluorescence in order to assess the degree of digestion of the ECM by the enzyme treatments.
Zaher M., Yelin R., Arraf A.A., Jadon J., Asleh M.A., Goltzman S., Shaulov L., Reinhardt D.P., & Schultheiss T.M. (2024). Stored Elastic Bending Tension as a Mediator of Embryonic Body Folding.
Publication 2024
2
Quantifying Calcium-Induced Cell Fragmentation
A-431-S cells were seeded into a 24-well plate and grown to confluence. Each monolayer was subjected to a 45 min calcium-free pulse and calcium was restored for 2, 4, 6, or 8 h, as described above. The no-pulse control was maintained in normal calcium throughout the experiment, while calcium was not restored for the time 0 control. After the calcium-free pulse assay, 1 U/mL Dispase II (Sigma-Aldrich) was added to each well for approximately 30 min – until the monolayer was entirely lifted from the plate. A P1000 tip was clipped and lubricated with FBS before subjecting each cell sheet to mechanical stress using a uniform number of pipette aspirations. The number of aspirations for each independent experiment was determined by the number required to produce countable fragments in the no-pulse control. After fragmentation, the samples were fixed with 1% paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA) and fragments were manually counted under a dissection microscope. The assay was performed independently three times, with each experiment comprising three technical replicates per time point.
Dean W.F., Albert R.M., Nawara T.J., Ubil M., Beggs R.R, & Mattheyses A.L. (2024). Dsg2 ectodomain organization increases throughout desmosome assembly. Cell Adhesion & Migration, 18(1), 1-13.
Full text: Click here
Publication 2024
3
Murine Lung Cell Isolation Protocol
Adult C57BL/6 mice over six weeks of age were anaesthetized with a 1:1 mixture of ketamine and medetomidine through intraperitoneal (IP) injection with a dose dependent on the body weight. Cervical dislocation was carried out, and death confirmed by terminal exsanguination. The abdominal aorta was cut, and the trachea exposed by removing the salivary glands. The trachea was then cannulated using a blunt needle and bronchoalveolar lavage with pre-warmed Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12) (Gibco) was performed (three times 0.8ml per mouse). The rib cage was then removed, and cardiovascular system was flushed by injecting 10ml of ice-cold DMEM/F12 into the right ventricle. A volume of 1.5-2ml of enzyme mix was then slowly injected through the tracheal cannula into the lungs, which were then re-moved (lungs were dissected out of the thoracic cavity by cutting at the primary bronchial bifurcation) and placed in 3ml of digestion mixture (2mg/ml Dispase II (Sigma-Aldrich) + 0.1mg/ml DNaseI (Sigma-Aldrich) re-suspended in DMEM/F12 (Gibco) + 1% v/v Penicillin-Streptomycin (Gibco)). The digestion mixture was previously filtered through 0.22μm filter and frozen. Lungs were then incubated for 1h at 37°C or 20h at 4°C. The digestion solution together with lungs was then poured onto a 70μm strainer, lungs were then dissociated using a 5ml syringe plunger by gently agitating the lungs on the mesh. The strainer was then washed using 10ml of dispase wash (DMEM/F12 (Gibco) + 0.05mg/ml DNaseI (Sigma-Aldrich) + 1% v/v Penicillin-Streptomycin (Gibco)). Each sample was then centrifuged for 15min, 4°C, 130 relative centrifugal force (RCF). The supernatant was then removed and 2ml of ice-cold ACK red blood cell lysis (Gibco) was added to each tube, samples were swirled for 90s, and 5ml of MACS buffer (Phosphate-buffered saline (PBS)—no Mg2+ and Ca2+ + 0.5% bovine serum albumin (BSA) + 2mM ethylenediaminetetraacetic acid (EDTA)) + 1% v/v Penicillin/Streptomycin (10,000 U/ml, Gibco) was added. Samples were then passed through a 40μm strainer, washed with 5ml of MACS buffer and centrifuged for 5min, 4°C at 300 RCF. Each sample was then resuspended in 1ml of MACS buffer and blocked with 5μl of 0.5mg/ml anti-mouse CD16/32 antibody (BioLegend) ready for further MACS processing or flow cytometry staining.
Janas P.P., Chauché C., Shearer P., Perona-Wright G., McSorley H.J, & Schwarze J. (2024). Cold dispase digestion of murine lungs improves recovery and culture of airway epithelial cells. PLOS ONE, 19(1), e0297585.
Full text: Click here
Publication 2024
4
Isolation of Human Epidermal Keratinocytes
Not available on PMC !
Freshly excised skin tissues were immersed in 0.25% dispase II (Sigma Aldrich, USA) overnight at 4 °C. Cells were washed with Hank's balanced salt solution (HBSS) and immersed in 0.25% dispase II for 5 min at 37 °C. Dispase solution was then replaced with HBSS. The epidermal tissues were carefully separated. The keratinocytes were carefully scraped together. Mechanical stress was applied by an electric pipette (Eppendorf, Germany) of 1 mL pipetting up and down for 5 times. The fragments of epidermis tissues and keratinocyte monolayers were counted.
Lai X., Wang M., Zhang Z., Chen S., Tan X., Liu W., Liang H., Li L, & Shao L. (2024). ZNPs reduce epidermal mechanical strain resistance by promoting desmosomal cadherin endocytosis via mTORC1-TFEB-BLOC1S3 axis. Journal of nanobiotechnology, 22(1).
Publication 2024
5
Pancreatic Single-Cell Isolation Protocols
For the dataset GSE181276, the pancreatic tissues were digested with the collagenase IV-dispase II (2 mg/ml) at 37 °C for 45 min and further treated with 0.25% trypsin solution at 37 °C for 5 min to obtain single-cell suspension [5 (link)]. For the dataset GSE188819, the pancreatic tissues were incubated with the mixture of digestive enzymes (1 mg/ml collagenase P, 2 U/ml dispase II, 0.1 mg/ml soybean trypsin inhibitor and 0.1 mg/ml DNase I in HBSS with Ca2+/Mg2+) and digested by a gentle MACS™ Octo Dissociator at 37 °C for 40 min, followed by further treatment with 0.05% Trypsin–EDTA at 37 °C for 5 min and removal of red blood cells [6 (link)].
Yang K., Xie R., Xiao G., Zhao Z., Ding M., Lin T., Tsang Y.S., Chen Y., Xu D, & Fei J. (2024). The integration of single-cell and bulk RNA-seq atlas reveals ERS-mediated acinar cell damage in acute pancreatitis. Journal of Translational Medicine, 22, 346.
Full text: Click here
Publication 2024

Top products related to «Dispase»

1
Dispase by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, Japan, Switzerland, France, Canada, Italy, China, Belgium
Dispase is an enzymatic cell dissociation reagent used for the isolation and dissociation of cells from various tissue types, including epithelial, endothelial, and connective tissues. It functions by breaking down extracellular matrix proteins, allowing for the efficient release of cells from their surrounding matrix.
2
Dispase 2 by Roche
Sourced in United States, Germany, Switzerland, Japan, United Kingdom, China, Austria, Israel, Canada, France
Dispase II is a highly purified protease enzyme that can be used for the dissociation of a variety of cell types, including epithelial, endothelial, and neural cells. It is an effective tool for the isolation and culture of primary cells from tissues.
3
Fetal bovine serum (fbs) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
4
Dispase 2 by Merck Group
Sourced in United States, Germany, United Kingdom, France, Switzerland, Israel, Canada
Dispase II is a proteolytic enzyme that can be used for the dissociation and isolation of cells from a variety of tissues. It is a neutral metalloprotease that effectively disperses cells while maintaining their viability and functionality.
5
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.
6
Dispase by Merck Group
Sourced in United States, Germany, Sao Tome and Principe, Switzerland, China, Japan, Israel, United Kingdom
Dispase is an enzyme primarily used for tissue dissociation and cell isolation. It is effective in disrupting the extracellular matrix, enabling the separation of cells from connective tissues.
7
Dispase by Roche
Sourced in United States, Germany, Switzerland, Japan, United Kingdom, China, France
Dispase is a neutral protease enzyme derived from Bacillus polymyxa. It is commonly used in cell and tissue dissociation applications to facilitate the isolation of cells from various tissue types.
8
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.
9
Penicillin streptomycin by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom, China, Canada, France, Japan, Australia, Switzerland, Israel, Italy, Belgium, Austria, Spain, Gabon, Ireland, New Zealand, Sweden, Netherlands, Denmark, Brazil, Macao, India, Singapore, Poland, Argentina, Cameroon, Uruguay, Morocco, Panama, Colombia, Holy See (Vatican City State), Hungary, Norway, Portugal, Mexico, Thailand, Palestine, State of, Finland, Moldova, Republic of, Jamaica, Czechia
Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
10
Collagenase dispase by Roche
Sourced in Switzerland, United States, Germany, United Kingdom, Japan
Collagenase/dispase is a laboratory enzyme product used for the dissociation and disaggregation of various tissue types, including soft connective tissues. It functions by breaking down the extracellular matrix components, such as collagen and other glycoproteins, to facilitate the isolation and harvesting of individual cells from the tissue.

More about "Dispase"

Dispase is a versatile proteolytic enzyme that has become an invaluable tool in cell and tissue dissociation procedures, particularly in the fields of stem cell research, tissue engineering, and regenerative medicine.
This enzyme, derived from the bacterium Bacillus polymyxa, has the remarkable ability to cleave a wide range of extracellular matrix components, making it highly effective in isolating cells from various tissues.
One of the key advantages of Dispase is its gentler approach compared to other dissociation enzymes, such as Collagenase or Trypsin.
This gentle action helps maintain the viability and functionality of the isolated cells, a crucial factor in downstream applications.
Researchers have also employed Dispase II, a variant of the original enzyme, which can provide additional benefits in specific experimental settings.
Beyond Dispase, the use of supporting agents like Fetal Bovine Serum (FBS), DNase I, and Penicillin/Streptomycin can further enhance the efficacy and reproducibility of cell dissociation protocols.
These supplementary components play important roles in minimizing cell clumping, preventing DNA-mediated cell aggregation, and maintaining a sterile environment.
When working with Dispase, it is essential to be aware of the potential for variability in its activity and effectiveness, which can be influenced by factors such as the source, lot, and experimental conditions.
Optimizing Dispase protocols through careful comparison and testing can help researchers achieve greater reproducibility and unlock new insights in their Dispase-based investigations.
Utilizing AI-driven platforms like PubCompare.ai can further enhance the efficiency and accuracy of Dispase research.
These tools can assist researchers in locating the best protocols from literature, preprints, and patents, while enabling seamless comparisons to identify the optimal solution for their specific needs.
By streamlining the Dispase experimentation process, researchers can unlock new possibilities and drive progress in the exciting realms of stem cell research, tissue engineering, and regenerative medicine.