Dispase type 2

Manufactured by Worthington

Sourced in United States

Dispase type II is a neutral protease enzyme derived from Bacillus polymyxa. It is used for the dissociation and isolation of cells from a variety of tissue types.

Automatically generated - may contain errors

Lab products found in correlation

Papain, by Worthington (7 mentions) Collagenase type 2, by Worthington (6 mentions) 1 cysteine, by Worthington (2 mentions) F12 media, by Thermo Fisher Scientific (2 mentions) Hsc 3, by American Type Culture Collection (1 mentions) Laminin, by Merck Group (1 mentions) B27 supplement, by Thermo Fisher Scientific (1 mentions) Percoll gradient, by Merck Group (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Poly d lysine, by Merck Group (1 mentions) Collagenase type 2 cls2, by Worthington (1 mentions)

7 protocols using dispase type 2

Culturing Cancer Cells and Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cancer cells: The human oral squamous cell carcinoma cell lines, HSC-3 and Hela-O3, were cultured in Dulbecco’s Modification of Eagle’s Medium (DMEM) with 4.5 g/L glucose, l-glutamine and sodium pyruvate, supplemented with 10% fetal bovine serum (FBS), and cultured at 37 °C in 5% CO₂. HSC-3 was purchased from ATCC and used fewer than 6 months after resuscitation. Hela-O3 was obtained from Dr. Roberto Weigert at the National Cancer Institute and tested for Mycoplasma by PCR (ATCC) prior to use.
Neurons: Mouse trigeminal ganglia were harvested and cultured as previously described^{26 (link)}. Trigeminal ganglia were isolated, transferred into Hank’s Balanced Salt Solution (HBSS) and enzyme-digested by incubation with papain (Worthington), collagenase type II (Worthington), and dispase type II (MB). Dissociated neurons were plated on glass coverslips coated with poly-d-lysine and laminin and maintained at 37 °C at 5% CO₂/95% air in F12 media (Life Technologies) with 5% FBS.

Dang D., Ye Y., Aouizerat B.E., Patel Y.K., Viet D.T., Chan K.C., Ono K., Doan C., Figueroa J.D., Yu G, & Viet C.T. (2020). Targeting the endothelin axis as a therapeutic strategy for oral cancer metastasis and pain. Scientific Reports, 10, 20832.

+ Open protocol

+ Expand

Isolation and Culture of Murine Trigeminal Ganglion Neurons

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Plates were coated with 20 μg/ml poly-D-Lysine (Sigma-Aldrich, St. Louis, United States) and 18 μg/ml Laminin (Sigma-Aldrich, St. Louis, United States) 1 day before neuron isolation, followed by overnight incubation at 4°C. As previously described (Katzenell et al., 2017 (link)), TGs were obtained from 6 weeks old C57B/6 mice and were digested first with 40 U/ml papain (Worthington, Lakewood, NJ, United States) in HBSS at 37°C for 20 min and then with 140 U/ml collagenase type II (Worthington, Lakewood, NJ, United States) and 0.4 U/ml dispase type II (Worthington, Lakewood, NJ, United States) in HBSS at 37°C for 20 min. Gradient separation was performed at 1300 × g for 10 min in the Percoll gradient (Sigma-Aldrich, St. Louis, United States), with slow acceleration and deceleration. Next, the isolated TG Neurons were inoculated onto a well-coated plate and incubated in Neurobasal-A Complete (NBA-C): Neurobasal A medium (Gibco, Grand Island, NY, United States) supplemented with 2% B-27 Supplements (Gibco, Grand Island, NY, United States), 1% GlutaMAX (Gibco, Grand Island, NY, United States). After 3–4 days of culture, the cells were used for follow-up experiments. A diagram of the process is shown in Figure 1.

Wang C., Liang Q., Sun D., He Y., Jiang J., Guo R., Malla T., Hamrah P., Liu X., Huang Z, & Hu K. (2022). Nectin-1 and Non-muscle Myosin Heavy Chain-IIB: Major Mediators of Herpes Simplex Virus-1 Entry Into Corneal Nerves. Frontiers in Microbiology, 13, 830699.

+ Open protocol

+ Expand

Acute Dissociation of Dorsal Root Ganglia Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse spinal columns were removed and placed in ice-cold HBSS; neurons were acutely dissociated and maintained as described ^E3,4. In brief, laminectomies were performed and bilateral DRG were dissected out. After removal of connective tissues, DRG were transferred to a 1 mL Ca²⁺/Mg²⁺-free HBSS containing 2 μL saturated NaHCO₃, 0.35 mg 1-cysteine and 20 U papain (Worthington, Lakewood, NJ, USA), and incubated at 37°C for 10 min. The suspension of DRG was centrifuged, the supernatant was removed, 1 mL Ca²⁺/Mg²⁺-free HBSS containing 4 mg collagenase type II and 1.25 mg dispase type II (Worthington) was added and incubated at 37°C for 10 min. After digestion, neurons were pelleted, suspended in neurobasal medium containing 2% B-27 supplement, 1% L-glutamine, l00 U·mL⁻¹ penicillin plus 100 μg·mL⁻¹ streptomycin, and 50 ng·mL⁻¹ nerve growth factor, plated on a 12 mm coverslip coated with poly-L-lysine (10 μg·mL⁻¹) and cultured under a humidified atmosphere of 5% CO₂/95% air at 37°C for 18–24 hr before use.

Feng J., Luo J., Mack M.R., Yang P., Zhang F., Wang G., Gong X., Cai T., Mei Z., Kim B.S., Yin S, & Hu H. (2017). The antimicrobial peptide hBD2 promotes itch through Toll-like receptor 4 signaling in mice. The Journal of allergy and clinical immunology, 140(3), 885-888.e6.

+ Open protocol

+ Expand

Acute Dissociation of Mouse DRG Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse spinal columns were removed and placed in ice-cold HBSS; neurons were acutely dissociated and maintained as described.^{13 (link)} In brief, laminectomies were performed and bilateral DRG were dissected out. After removal of connective tissues, DRG were transferred to a 1 mL Ca²⁺/Mg²⁺-free HBSS containing 2 μL saturated NaIICO₃, 0.35 mg 1-cysteine and 20 U papain (Worthington, Lakewood, NJ, USA), and incubated at 37°C for 10 min. The suspension of DRG was centrifuged, the supernatant was removed, 1 mL Ca²⁺/Mg²⁺-free HBSS containing 4 mg collagenase type II and 1.25 mg dispase type II (Worthington) was added and incubated at 37°C for 10 min. After digestion, neurons were pelleted, suspended in neurobasal medium containing 2% B-27 supplement, 1% 1-glutamine, 100U·mL⁻¹ penicillin plus 100μg·mL⁻¹ streptomycin, and 50ng·mL⁻¹ nerve growth factor, plated on a 12mm coverslip coated with poly-l-lysine (10 μg·mL⁻¹) and cultured under a humidified atmosphere of 5% CO₂/95% air at 37°C for 18–24 hr before use.

Luo J., Feng J., Yu G., Yang P., Mack M.R., Du J., Yu W., Qian A., Zhang Y., Liu S., Yin S., Xu A., Cheng J., Liu Q., O’Neil R.G., Xia Y., Ma L., Carlton S.M., Kim B.S., Renner K., Liu Q, & Hu H. (2017). TRPV4-expressing Macrophages and Keratinocytes Contribute Differentially to Allergic and Non-allergic Chronic Itch. The Journal of allergy and clinical immunology, 141(2), 608-619.e7.

+ Open protocol

+ Expand

Isolation and Culture of Mouse TG Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse TG neurons were harvested and cultured as previously described [27 (link)]. Briefly, BALB/c mice were euthanized with isoflurane. Trigeminal ganglia were removed, transferred into HBSS and enzyme-digested by incubation with papain (Worthington), collagenase type II (CLS2) (Worthington), and dispase type II (MB). Dissociated neurons were plated on glass coverslips coated with poly-d-lysine and laminin and maintained for approximately 2 hr at 37°C at 5% CO₂/95% air in F12 media (Gibco BRL) supplemented with 10% FBS.

Ye Y., Ono K., Bernabé D.G., Viet C.T., Pickering V., Dolan J.C., Hardt M., Ford A.P, & Schmidt B.L. (2014). Adenosine triphosphate drives head and neck cancer pain through P2X2/3 heterotrimers. Acta Neuropathologica Communications, 2, 62.

+ Open protocol

+ Expand

Isolation and Culture of Dorsal Root Ganglia Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were killed by cervical dislocation following CO₂ asphyxia and spinal columns were removed and placed in ice-cold HBSS. Laminectomies were performed and bilateral DRGs were dissected out. After removal of connective tissues, DRGs were digested in 1 mL of Ca²⁺/Mg²⁺-free HBSS containing 20 U of papain (Worthington, Lakewood, NJ), 0.35 mg of l-cysteine and 1 μL of saturated NaHCO₃ and incubated at 37 °C for 10 min. The DRG suspension was centrifuged, the supernatant was removed, and 1 mL of Ca²⁺/Mg²⁺-free HBSS containing 4 mg of collagenase type II and 1.25 mg of Dispase type II (Worthington) was added and incubated at 37 °C for 15 min. After digestion, neurons were pelleted; suspended in neurobasal medium containing 1% l-glutamine, 2% B-27 supplement, 100 U mL⁻¹ penicillin plus 100 μg mL⁻¹ streptomycin, and 50 ng mL⁻¹ nerve growth factor. The cells were plated on a 12-mm coverslip coated with poly-l-lysine (10 μg mL⁻¹) and cultured under a humidified atmosphere of 5% CO₂/95% air at 37 °C for 24 h.

Arnold W.R., Carnevale L.N., Xie Z., Baylon J.L., Tajkhorshid E., Hu H, & Das A. (2021). Anti-inflammatory dopamine- and serotonin-based endocannabinoid epoxides reciprocally regulate cannabinoid receptors and the TRPV1 channel. Nature Communications, 12, 926.

+ Open protocol

+ Expand

Culturing Dorsal Root Ganglion Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Dorsal root ganglion (DRG) neurons were primarily cultured, as described previously. Briefly, thoracic and lumbar DRGs were dissected and collected from adult mice and transferred into HBSS without Ca²⁺/Mg²⁺ on ice. F12 with 10% FCS and 100 U/ml penicillin/streptomycin was used as a culture medium. Ganglia were incubated with 1.5 mL papain (40 U/ml, Worthington) for 10 min at 37°C and then with 3 mL collagenase type II (4 mg/ml, Worthington)/ dispase type II (4.67 mg/ml, Worthington) combined solution for 10 min at 37°C. Dissociated cells were plated on poly-D-lysine-treated small coverslips and incubated for 2 to 3 days at 37°C in 95% air/5% CO².

Xu J., Zanvit P., Hu L., Tseng P.Y., Liu N., Wang F., Liu O., Zhang D., Jin W., Guo N., Han Y., Yin J., Cain A., Hoon M.A., Wang S, & Chen W. (2020). The Cytokine TGF-β Induces Interleukin-31 Expression from Dermal Dendritic Cells to Activate Sensory Neurons and Stimulate Wound Itching. Immunity, 53(2), 371-383.e5.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!