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Our neuron dissociation solutions can be used to dissociate and isolate neurons from tissues derived from the central nervous system of rat and mouse. It is composed of 3 solutions (enzyme, dissociation, and isolation solutions).
The product is ready-to-use without the need for preparations, and achieves simple isolation of neurons while maintaining high cell viability.
4. Washing solution: Leibovitz’s L-15 medium (code No. 128-06075) or HPSS (+) (code No.128-06075)
| Experimental condition | Type of solution | Amount | Recommended products |
|---|---|---|---|
| For samples containing large tissues that need to be dissociated (1-3 cerebral hemispheres from mouse or rat embryos) |
Enzyme solution | 5.0mL | Neuron Dissociation Solutions (Code: 291-78001) |
| Dispersion solution | 3.0mL | ||
| Isolation solution | 5.0mL | ||
| For samples containing small tissues that need to be dissociated (1-8 hippocampi from mouse or rat embryos) |
Enzyme solution | 2.5mL | Neuron Dissociation Solutions S (Code: 297-78101) |
| Dispersion solution | 2.5mL | ||
| Isolation solution | 2.5mL |
Frozen neurons from our company were thawed, and were dissociated and isolated in the neuron dissociation solutions.
| Product Name | |||
|---|---|---|---|
| ① | Hippocampus, from mouse (embryonic day 16) | Total(cells/vial) | 4.32 x 105 |
| Viable(cells/vial) | 3.83 x 105 | ||
| Cell viability(%) | 89 | ||
| ② | Cerebral cortex, from mouse (embryonic day 15) | Total(cells/vial) | 6.64 x 106 |
| Viable(cells/vial) | 6.09 x 106 | ||
| Cell viability(%) | 92 | ||
| ③ | Hippocampus, from rat (embryonic day 19) | Total(cells/vial) | 1.37 x 106 |
| Viable(cells/vial) | 1.24 x 106 | ||
| Cell viability(%) | 90 | ||
| ④ | Cerebral cortex, from rat (embryonic day 17) | Total(cells/vial) | 1.32 x 107 |
| Vialble(cells/vial) | 1.17 x 107 | ||
| Cell viability(%) | 89 | ||
| ⑤ | Cerebral striatum, from rat (embryonic day 17) | Total(cells/vial) | 2.73 x 106 |
| Viable(cells/vial) | 2.58 x 106 | ||
| Cell viability(%) | 95 |
▶The data demonstrated that the use of neuron dissociation solutions enables isolation of cells from frozen samples with high viability (~90%).
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NS supplement is a serum-free supplement for culturing neurons. It can be used to culture neurons and neural stem cells isolated from rat hippocampus. It should be used with NS basal medium and 200 mmol/L L-glutamine solution. Vitamin A-free and insulin-free products are also available.
NS supplement
Serum-free supplement for culturing neurons
NS supplement without insulin
NS supplement that does not contain insulin
Suitable for use in studying insulin secretion and receptors
NS supplement without vitamin A
NS supplement that does not contain vitamin A (retinyl acetate)
Vitamin A is considered to be involved in the differentiation of cells into neurons; thus, vitamin A-free reagents are used especially for culturing neural stem cells and glial cells.
Figure 1: Comparison of the number of cells
Neurons were isolated from the hippocampus of rat embryo (E19), and were cultured on a plate coated with poly-L-lysine. NS supplement was added to NS basal medium to the final concentration of 2%. Neurons were cultured for 5 days and the number of cells were compared.
Green: neuron-specific marker (TuJ1) Blue: nucleus (DAPI)
Blue: nucleus (DAPI)
Figure 2: Expression of a neuron-specific marker
Cells were stained with a neuron-specific marker (TuJ1) and a nuclear marker (DAPI).
Neurons were isolated from the cerebral cortex of rat embryo (E17), and were cultured in NS basal medium supplemented with either insulin-free or insulin-containing NS supplement (NSS). Cells were cultured for 6 days, and the number of cells grown with insulin-free supplement was compared relative to the number of cells grown with NSS (=1). As a comparison, cells were also cultured with insulin-free supplement containing 0.25x and 1.0x the amount of insulin in NSS.
Culture medium:
NS basal medium + 2% NS supplement without insulin or NSS + 0.5 mmol/L L-glutamine (+ 0.25x or 1.0x insulin)
Number of cells seeded:
4.0×104 cells/well(96-well plate coated with poly-L-lysine (30-70 kDa)
Since insulin promotes survival of neurons, the number of viable cells was lower when they were grown with insulin-free NS supplement. However, with the addition of insulin, the number of cells was equivalent to those grown with NSS.
Neural stem cells were isolated from rat hippocampus, and were cultured in NS basal medium supplemented with either vitamin A-free or vitamin A-containing NS supplement (NSS). Cells were cultured for 3 days, and the number of cells grown with vitamin A-free supplement was compared relative to the number of cells grown with NSS (=1).
< Culture medium >
NS basal medium + 2% NS supplement without vitamin A, Competitor’s product, or NSS + 1 mmol/L L-glutamine
< Number of cells seeded >
12,000 cells/well (96-well plate coated with poly-L-lysine)
The numbers of cells grown with our NS supplement without vitamin A and the equivalent product from another company were comparable.
Neurons were isolated from the hippocampus of rat embryo (E19), and were cultured in NS basal medium supplemented with either our vitamin A-free NS supplement or an equivalent vitamin A-free product from another company. Cells were cultured for 6 days, and the expression of neuron-specific marker (β-III-tubulin, TuJ1) was examined.
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NS Basal Medium is a chemically-defined synthetic medium optimized for culturing nerve cells. NS Supplement is a serum-free supplement (containing retinyl acetate) specifically designed for supporting the growth of nerve cells. When mixed together, they can be used for culturing a variety of nerve cells, including rat hippocampal neurons and neural stem cells.
Rat hippocampal neurons isolated from E19 embryos were cultured using poly-L-lysine-coated microtiter plates.
The morphology of these neurons was analyzed on day 6. We also examined the expression of the neuronal markers MAP2(a+b) and the glial marker GFAP on day 21
< Growth Medium >
NS Basal Medium containing 2% NS Supplement and 0.5 mM L-glutamine
< Seeding Density >
13,000cells/well（96well plate）
Primary cortical neurons isolated from E17 rat embryos were cultured using poly-L-lysine-coated microtiter plates.
The morphology of these neurons was analyzed on day 8. We also examined the expression of the neuronal markers MAP2(a+b) and the glial marker GFAP on day 14.
< Growth Medium >
NS Basal Medium containing 2% NS Supplement and 0.5 mM L-glutamine
< Seeding Density >
60,000cells/well（96well plate）
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The products in this series are frozen brain tissues collected from rat and mouse embryos. They reduce the time needed to prepare the animal models. Use our neuron dissociation solution (code No. 291-78001,297-78101) to dissociate the tissues, and use our neuron culture medium (code No. 148-09671) to culture cells collected from the tissues.
The products in our frozen neuron series were thawed, and our dissociation solution was used to disperse and isolate the cells. Cell viability was calculated as % of total cells.
| Product name | |||
|---|---|---|---|
| ① | Hippocampus, from mouse (embryonic day 16) |
Total number of cells(cells/vial) | 4.32 x 105 |
| Total number of viable cells(cells/vial) | 3.83 x 105 | ||
| Cell viability(%) | 89 | ||
| ② | Cerebral cortex, from mouse (embryonic day 15) |
Total number of cells(cells/vial) | 6.64 x 106 |
| Total number of viable cells(cells/vial) | 6.09 x 106 | ||
| Cell viability(%) | 92 | ||
| ③ | Hippocampus, from rat (embryonic day 19) |
Total number of cells(cells/vial) | 1.37 x 106 |
| Total number of viable cells(cells/vial) | 1.24 x 106 | ||
| Cell viability(%) | 90 | ||
| ④ | Cerebral cortex, from rat (embryonic day 17) |
Total number of cells(cells/vial) | 1.32 x 107 |
| Total number of viable cells(cells/vial) | 1.17 x 107 | ||
| Cell viability(%) | 89 | ||
| ⑤ | Cerebral striatum, from rat (embryonic day 17) |
Total number of cells(cells/vial) | 2.73 x 106 |
| Total number of viable cells(cells/vial) | 2.58 x 106 | ||
| Cell viability(%) | 95 |
The data demonstrated that neurons were isolated from frozen samples with approximately 90% viability.
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StemSure® Gelatin Solution is a coating agent that can be used for cell culture dishes and multi-well plates.
It is composed of gelatin derived from porcine skin, and is prepared with D-PBS (-).
Multi-well plates were coated with StemSure® Gelatin Solution, and feeder sells were seeded onto the coated wells. Human iPS cell line 201B7 was cultured on the feeder cells, and the morphology of the iPS cells and positivity for ALP staining were examined.
D-MEM/Ham’s F-12 + 20% StemSure® Serum Replacement + 2 mmol/L L-Glutamine + 1 x MEM Non-essential Amino Acids + 0.1 mmol/L StemSure® 2-Mercaptoethanol +1 x Penicillin-Streptomycin + 5 ng/mL bFGF
Cloning efficiency for murine ES cell line D3 was compared for cells cultured on multi-well plates coated with StemSure® Gelatin Solution and those cultured on plates coated with an equivalent material from company A.
Murine ES cell line D3 was cultured on a multi-well plate coated with StemSure® Gelatin Solution. After 16 passages, cells stained positive for markers of undifferentiated cells (Oct3/4, Sox2, Nanog).
Murine ES cell line D3 was cultured for multiple passages on multi-well plates coated with StemSure® Gelatin Solution. Cells were then injected subcutaneously in immunocompromised mice. Teratomas formed subcutaneously, and various tissue types including nerve tissue (derived from ectoderm), cartilage tissue (derived from mesoderm), and luminal structures with ciliated epithelium (derived from endoderm) were found in the teratomas.
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Vitronectin is a 478 amino-acid glycoprotein found in serum and the extracellular matrix. Similar to other extracellular matrix proteins (such as fibronectin and laminins), it is classified as a cell adhesion protein. Vitronectin is multifunctional; in addition to promoting cell adhesion and spreading, it plays a role in both the complement and coagulation cascades.
FUJIFILM Wako’s vitronectin is a recombinant protein. It lacks the N-terminal signal peptide and thus contains amino-acid residues 20-398. This product is ideal for functional studies of vitronectin in the areas of tissue repair and regeneration. It can also be used as a chemically-defined biomaterial for various cell culture applications, including scaffold construction.
Human iPS cells were passaged six times using 6-well plates coated with this product. The maintenance of the undifferentiated state in these cells was confirmed by the expression of the indicated pluripotency markers.
Cell Line: hiPS201B7
Medium: StemSure®® hPSC medium Δ (Product Code: 197-17571) containing 32 ng/mL bFGF (Product Code: 060-05383)
< Product Information >
| Purity | ≥90% (SDS-PAGE) |
| Expression system | E. coli |
| Form | Liquid |
| Concentration | 0.5mg/mL |
| Formulation | 20mM Tris-HCl, pH8.0 (Contains NaCl, KCl, EDTA, arginine, DTT, and glycerol) |
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Learn more for the detailed instructions and protocol
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Nippi Inc. provides animal collagen products for research use.
◆Collagen type I
Type I collagen is the major fibrous protein in the extracellular matrix. It can be found in most organs, but is particularly abundant in bone, tendon, and dermis.
Acid Soluble Collagen(ASC)
・Due to its high gel strength, this collagen preparation is ideal for both “On-Gel” and “In-Gel (Gel-Embedded)” cultures.
・Filtered through a 0.45 μm pore size membrane.
Pepsin Solubilized Collagen (PSC)
・ Atelocollagen prepared by enzymatically removing telopeptides.
・ This form of collagen has high cellular adhesiveness, making it perfect for coating cell culture vessel surfaces.
・ Filtered through a 0.45 μm pore size membrane.
◆Collagen type Ⅲ
This is a fibrillar collagen abundant in tissues with high elasticity, such as juvenile skin tissues and the arterial tunica media where smooth muscle cells populate. Type III collagen is also expressed during embryogenesis and wound healing.
・Atelocollagen prepared by enzymatically removing telopeptides.
・Filtered through a 0.45 μm pore size membrane.
◆Collagen type Ⅳ
Type IV collagen is one of the major components of basement membranes. It polymerizes to form a mesh-like network in the basal lamina.
・Acid-extracted native collagen.
・This form of collagen has high cellular adhesiveness . It is suitable for collagen-gel cultures and collagen-coating of culture vessel surfaces.
Collagen has long been available only as either a liquid or lyophilized preparation. Nippi Inc. used its proprietary technology and succeeded in producing easy-to-use collagen powder that maintains the protein’s native structure (triple helix). (International patent pending.) You can now prepare a collagen solution at a desired concentration using the solvent of your choice.
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Nitta Gelatin’s collagen- and gelatin-based reagents are trusted products with proven performance that have long been used by many researchers. Their collagen products have superior optical clarity even after gel formation is completed. This provides an opportunity to accurately observe biological samples embedded in collagen gel matrices.
| ●Type Ⅰ-A ・Ideal for collagen-gel cultures. ・Perfect for microscopy due to its outstanding transparency. |
●Type Ⅰ-P ・Recommended for both collagen-gel cultures and collagen-coating of culture surfaces. ・Forms softer gels than Type I-A. ・Pepsin-solubilized collagen from porcine tendons. |
| ●Type Ⅰ-C ・Recommended for collagen-coating of culture vessels surfaces. ・Pepsin-solubilized collagen from porcine skin. |
●Type Ⅲ ・Recommended for collagen-coating of culture vessels surfaces. ・Does not form gels. ・Type-III collagen from porcine skin. |
| ●Type Ⅳ ・Recommended for collagen-coating of culture surfaces. ・Does not form gels. ・Type-IV collagen; purified from bovine anterior lens capsules using pepsin treatment. |
When you grow your cells…
The Collagen-Gel Cell Culturing Kit is the right choice for researchers interested in implementing collagen cultures (2D/3D) using Cellmatrix® I-A. This convenient starter kit contains all the reagents necessary for preparing collagen gel matrices.
Specifically prepared for collagen-gel cultures
Available Media Types
o 10x Concentrates: Ham’s F-12 medium, MEM with Hanks’ balanced salt solution, DF medium (DME:F-12 = 1:1), and Medium 199.
o 5x Concentrates: DME medium and RPMI-1640.
Specifically formulated for collagen-gel cultures.
For researchers and industry professionals seeking higher levels of safety and quality in their materials for biomedical engineering and clinical applications.
Virus-inactivated collagen and gelatin with exceptionally low levels of endotoxins.
Endotoxin Level: ≤10 EU/g
Nitta Gelatin Inc. has submitted a Device Master File (DMF) with the United States FDA for beMatrix® Gelatin products.
MedGel® Ⅱ is an improvement on the original MedGel® (“TaBaTa Gel for controlled release” outside of Japan),
a bioabsorbable gelatin hydrogel for the sustained release of physiologically active substances.
[Applications]
・Drug delivery research and the development of new drug delivery systems (DDSs).
・Basic and clinical research in cell therapy and regenerative medicine.
[Improvements on the Original MedGel®]
・Changed the method of sterilization to γ irradiation.
・Implemented a new crosslinking method that eliminates the need for chemical crosslinkers.
MedGel® (“TaBaTa Gel for controlled release” outside of Japan) is a bioabsorbable gelatin based hydrogel that can release physiologically active substances in a gradual and continuous manner. It was developed based on research by Prof. Yasuhiko Tabata of the Institute for Frontier Medical Sciences at Kyoto University. MedGel® is a water-insoluble polymer network formedby crosslinking gelatin. This polymeric network captures therapeutic agents primarily via electrostatic interactions. Following in vivo implantation MedGel® – therapeutic reagent complex, MedGel® is degraded by endogenous enzymes, allowing entrapped drugs to diffuse out of the network. MedGel® is also designed to be completely degraded and absorbed in the body. This eliminates the need for implant removal surgery after the drugs are exhausted.
[Sheet Type]
Nitta Gelatin Inc. offers two products (PI5 and PI9) with different isoelectric points. This enables you to select the hydrogel suitable for the isoelectric point (PI) property of your drug compound.
[Particle Type]
This form of hydrogel can be delivered via injection with a needle and syringe.
Gelatin from porcine skin (type A) or bovine bones (type B) for high-strength gels.
Applications
Features
Applications
Features
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