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can be used to investigate many aspects of neuronal biology

Mouse Schwann Cell Line (IMS32) Culture Kit

Background

Schwann cells, the principle glial cells of the peripheral nervous system (PNS), are involved in several aspects of PNS biology. In addition to providing trophic support for neuronal development and maintenance through the production of cytokines and growth factors, Schwann cells wrap around motor and sensory neuron axons to create an insulating myelin sheath that aids impulse conductivity. In recent years, Schwann cells have become increasingly recognized for their support to nerve regeneration while their dysfunction may contribute to neurodegenerative diseases such as ALS. IMS32 cells are a spontaneously immortalized cell line established from long term culture of mouse dorsal root ganglia and peripheral nerves (1,2). IMS32 ce lls retain many characteristic of mature Schwann cells and are readily propagated in the supplied media. IMS32 cells can be used to investigate many aspects of neuronal biology including nerve regeneration, neuroprotective mechanisms, and for the development of novel therapeutic approaches to neurological disorders.

Kit Components

*Shipping: dry ice
Note: Cosmo Bio only guarantees growth of this cell line in Cat No. PMC-SWN-MM-COS.
Product Name Quantity Amount Storage Stability
Mouse Schwann Cell Line(IMS32), Cryopreserved 5 x 105 cells/vial 1 liquid nitrogen, vapor phase 6 months
Mouse Schwann Cell Culture Medium (Cat No. PMC-SWN-MM-COS) Main components: D-MEM, FBS and antibiotic 500 mL 1 -20°C Freezer 6 months
4°C 3 months

Protocols

[Thawing Procedure]-For 100 mm plastic culture dish.

  1. Bring IMS32 Culture Media to room temperature.
  2. Put 10 mL of culture medium into a 50 mL conical tube.
  3. Remove cryovial from liquid nitrogen and immediately thaw cells in a 37°C water bath for 2 minutes.
    Caution: Do NOT vortex the cells
  4. Using a pipette, transfer thawed contents of cryovial to the 50 mL conical tube.
  5. Rinse the cryovial with 1 mL of cell suspension to collect remaining cells.
  6. Transfer the cell suspension to a 100 mm plastic culture dish (cell density will be ~9 x 104 cells/cm2) and incubate at 37°C under 5% CO2, 100% humidity.
    Note: Centrifugation of cells after thawing is not recommended. Centrifugation is more harmful to cells than the effect of residual cell freezing medium remaining in the culture following this procedure. Centrifugation at this time may lead to a low proliferation rate and poor cell attachment.
  7. Replace culture medium the next day to remove residual cell freezing medium and unattached cells, then every 2 - 3 days thereafter.
    Note: After 3~5 days, IMS32 cells will reach approximately 80~100% confluence.
    Note: IMS32 cells do not adhere to untreated glass. For culture on glass slides or glass dish, pre-coat culture surface with Collagen I or poly-L-lysine.
    NOTE: Cell attachment and proliferation on coated glass surfaces is generally weaker than on plastic.

[Subculturing Procedure]

  1. Use IMS32 cells at 70~90% confluence. See Image C below.
  2. Remove spent medium from the dish.
  3. Add 10~12 mL of pre-warmed IMS32 Culture Medium.
  4. Detach cells from dish by pipetting. Cells will detach easily. (Trypsin or other cell dissociation enzyme should not be used). Transfer cell suspension to a 50 mL centrifuge tube.
    Note: Use microscope to observe dish following detachment. If more than 10% of cells remain undetached, growth of subcultured cells will be slow.
  5. Split cells 1:3 or 1:4 with IMS32 Culture Medium into fresh cell culture dishes. Return to incubator.

[Freezing Procedure]

  1. Follow steps 1-4 [Subculturing Procedure] above.
  2. Centrifuge the 50 mL tube at ~200 × g for 5 minutes at room temperature. Remove supernatant without disturbing the pelleted cells.
  3. Resuspend cell pellet to ~1 × 106 cells/mL with cold COS Banker Cell Freezing Medium (Cosmo Bio Co., Ltd. Cat No. KOJ-COS-CFM01) or other standard DMSO-based freeze media.
  4. Dispense aliquots of this cell suspension into cryogenic storage vials. Keep vials in cell freezing container (e.g. CoolCell®, Biocision). Store vilas at -80°C overnight.
  5. Transfer the vials into gas phase above liquid nitrogen for long term storage.

Schwann Cell Line
A
Schwann Cell Line
B
Schwann Cell Line
C
Immunofl uorescence of IMS32 cells
A: Anti-p75NTR (green), nuclear staining (blue)
B: Anti-s100 (green), nuclear staining (blue)
C: phase-contrast microscopic image

Product List

Product Name Cat# Quantity Price

Schwann Cell Line (IMS32) Culture Kit

PMC-SWN-IMS32-COS 1KIT

¥ 85,000
$ 1134
€ 850

Schwann Cell (IMS32) Medium

PMC-SWN-MM-COS 500ML

¥ 27,500
$ 367
€ 275

Anti Schwann Cell/peripheral myelin

CAC-GU01-M01AS-A 50UL

¥ 30,000
$ 400
€ 300

References
  • 1. Spontaneously immortalized adult mouse Schwann cells secrete autocrine and paracrine growth-promoting activities. Watabe K, Fukuda T, Tanaka J, et al. J Neurosci Res. 1995;41:279-290. PMID: 7650763
  • 2. Tissue culture methods to study neurological disorders: Establishment of immortalized Schwann cells from murine disease models. Watabe K, Sakamoto T, Kawazoe Y, Michikawa M, Miyamoto K, Yamamura T, Saya H, Araki N. Neuropathology 2003;23:64-74. PMID: 1272292

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.