PMID- 29435762
OWN - NLM
STAT- MEDLINE
DCOM- 20180612
LR  - 20181113
IS  - 1432-119X (Electronic)
IS  - 0948-6143 (Linking)
VI  - 149
IP  - 5
DP  - 2018 May
TI  - Establishment of a myelinating co-culture system with a motor neuron-like cell 
      line NSC-34 and an adult rat Schwann cell line IFRS1.
PG  - 537-543
LID - 10.1007/s00418-018-1649-x [doi]
AB  - Co-culture models of neurons and Schwann cells have been utilized for the study 
      of myelination and demyelination in the peripheral nervous system; in most of the 
      previous studies, however, these cells were obtained by primary culture with 
      embryonic or neonatal animals. A spontaneously immortalized Schwann cell line 
      IFRS1 from long-term cultures of adult Fischer rat peripheral nerves has been 
      shown to retain fundamental ability to myelinate neurites in co-cultures with 
      adult rat dorsal root ganglion neurons and nerve growth factor-primed PC12 cells. 
      Our current investigation focuses on the establishment of stable co-culture 
      system with IFRS1 cells and NSC-34 motor neuron-like cells. NSC-34 cells were 
      seeded at a low density (2 x 10(3)/cm(2)) and maintained for 5-7 days in 
      serum-containing medium supplemented with non-essential amino acids and 
      brain-derived neurotrophic factor (BDNF; 10 ng/mL). Upon observation of neurite 
      outgrowth under a phase-contrast microscope, the NSC-34 cells were exposed to an 
      anti-mitotic agent mitomycin C (1 microg/mL) for 12-16 h, then co-cultured with IFRS1 
      cells (2 x 10(4)/cm(2)), and maintained in serum-containing medium supplemented 
      with ascorbic acid (50 microg/mL), BDNF (10 ng/mL), and ciliary neurotrophic factor 
      (10 ng/mL). Double immunofluorescence staining carried out at day 28 of the 
      co-culture showed myelin protein (P0 or PMP22)-immunoreactive IFRS1 cells 
      surrounding the betaIII tubulin-immunoreactive neurites. This co-culture system can 
      be a beneficial tool to study the pathogenesis of motor neuron diseases (e.g., 
      amyotrophic lateral sclerosis, Charcot-Marie-Tooth diseases, and immune-mediated 
      demyelinating neuropathies) and novel therapeutic approaches against them.
FAU - Takaku, Shizuka
AU  - Takaku S
AD  - Diabetic Neuropathy Project, Department of Sensory and Motor Systems, Tokyo 
      Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, 
      Tokyo, 156-8506, Japan.
FAU - Yako, Hideji
AU  - Yako H
AD  - Diabetic Neuropathy Project, Department of Sensory and Motor Systems, Tokyo 
      Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, 
      Tokyo, 156-8506, Japan.
FAU - Niimi, Naoko
AU  - Niimi N
AD  - Diabetic Neuropathy Project, Department of Sensory and Motor Systems, Tokyo 
      Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, 
      Tokyo, 156-8506, Japan.
FAU - Akamine, Tomoyo
AU  - Akamine T
AD  - Diabetic Neuropathy Project, Department of Sensory and Motor Systems, Tokyo 
      Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, 
      Tokyo, 156-8506, Japan.
AD  - Division of Diabetes, Metabolism and Endocrinology, Department of Internal 
      Medicine, Jikei University School of Medicine, Minato-ku, Tokyo, 105-8461, Japan.
FAU - Kawanami, Daiji
AU  - Kawanami D
AD  - Division of Diabetes, Metabolism and Endocrinology, Department of Internal 
      Medicine, Jikei University School of Medicine, Minato-ku, Tokyo, 105-8461, Japan.
FAU - Utsunomiya, Kazunori
AU  - Utsunomiya K
AD  - Division of Diabetes, Metabolism and Endocrinology, Department of Internal 
      Medicine, Jikei University School of Medicine, Minato-ku, Tokyo, 105-8461, Japan.
FAU - Sango, Kazunori
AU  - Sango K
AUID- ORCID: 0000-0002-9750-9596
AD  - Diabetic Neuropathy Project, Department of Sensory and Motor Systems, Tokyo 
      Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, 
      Tokyo, 156-8506, Japan. sango-kz@igakuken.or.jp.
LA  - eng
GR  - JSPS KAKENHI 16K07048/the Ministry of Education, Science, Sports and Culture of 
      Japan/
PT  - Journal Article
DEP - 20180212
PL  - Germany
TA  - Histochem Cell Biol
JT  - Histochemistry and cell biology
JID - 9506663
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Coculture Techniques/*methods
MH  - Motor Neurons/*cytology/*metabolism
MH  - Myelin Sheath/*metabolism
MH  - Rats
MH  - Schwann Cells/*cytology/*metabolism
OTO - NOTNLM
OT  - Co-culture
OT  - Immortalized Schwann cells
OT  - Mitomycin C
OT  - Motor neuron-like cells
OT  - Myelination
EDAT- 2018/02/13 06:00
MHDA- 2018/06/13 06:00
CRDT- 2018/02/14 06:00
PHST- 2018/02/06 00:00 [accepted]
PHST- 2018/02/13 06:00 [pubmed]
PHST- 2018/06/13 06:00 [medline]
PHST- 2018/02/14 06:00 [entrez]
AID - 10.1007/s00418-018-1649-x [pii]
AID - 10.1007/s00418-018-1649-x [doi]
PST - ppublish
SO  - Histochem Cell Biol. 2018 May;149(5):537-543. doi: 10.1007/s00418-018-1649-x. 
      Epub 2018 Feb 12.