Improved culture methods to expand Schwann cells with altered growth behaviour from CMT1A patients.
A duplication of the gene for myelin protein PMP22 is by far the most common cause of the hereditary demyelinating neuropathy CMT1A. A role for PMP22 in cell growth in addition to its function as a myelin protein has been suggested because PMP22 is homologous to a gene specifically upregulated during growth arrest. Furthermore, transfected rat Schwann cells overexpressing PMP22 show reduced growth. In addition, abnormal Schwann cell differentiation has been described in nerve biopsies from CMT1A patients. To analyse whether the duplication of the PMP22 gene in CMT1A neuropathy primarily alters Schwann cell differentiation and to exclude nonspecific secondary responses, we improved human Schwann cell culturing. This allowed us long-term passaging of human Schwann cells with unchanged phenotype, assessed by expression of different Schwann cell markers. Subsequently we established Schwann cell cultures from CMT1A nerve biopsies. We find decreased proliferation of Schwann cells from different CMT1A patients in all passages. We also demonstrate PMP22 mRNA overexpression in cultured CMT1A Schwann cells. We conclude that decreased proliferation in cultured Schwann cells that carry the CMT1A duplication indicates abnormal differentiation of CMT1A Schwann cells. The identification of an abnormal phenotype of CMT1A Schwann cells in culture could possibly lead to an in vitro disease model.
Hanemann
C O
CO
Department of Neurology, Heinrich-Heine-University, Duesseldorf, Germany. C.O.Hanemann@uni-duesseldorf.de
Rosenbaum
C
C
Kupfer
S
S
Wosch
S
S
Stoegbauer
F
F
Müller
H W
HW
D000941:Antigens / Q000032:analysis; D015415:Biomarkers; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002607:Charcot-Marie-Tooth Disease / Q000473:pathology*; D006801:Humans; D009185:Myelin Proteins / Q000032:analysis; D012583:Schwann Cells / Q000276:immunology / Q000473:pathology*
D000941:Antigens; D015415:Biomarkers; D009185:Myelin Proteins; C073687:PMP22 protein, human; C489546:Pmp22 protein, rat