Dopaminergic Differentiation of the Nurr1-expressing Immortalized Mesencephalic Cell Line CSM14.1 in Vitro

Overview

Journal J Anat

Specialty Anatomy & Histology

Date 2002 Aug 13

PMID 12171477

Citations 7

Authors

Stefan Jean-Pierre Haas

Andreas Wree

Affiliations

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Abstract

The use of neural stem cells as grafts is a potential treatment for Parkinson's disease, but the potential of stem cells to differentiate into dopaminergic neurones requires investigation. The present study examined the in vitro differentiation of the temperature-sensitive immortalized mesencephalic progenitor cell line CSM14.1 under defined conditions. Cells were derived from the mesencephalic region of a 14-day-old rat embryo, retrovirally immortalized with the Large T antigen and cultured at 33 degrees C in DMEM containing 10% fetal calf serum (FCS). For differentiation, the temperature was elevated at 39 degrees C and FCS was reduced (1%). Using histology, immunocytochemical detection of the stem cell marker Nestin and the neuronal marker MAP5 and, in addition, Western blotting to determine the presence of neurone-specific enolase and the neurone nuclei antigen we demonstrated a differentiation of these cells into neuronal cells accompanied by a decrease in Nestin production. In Western blots, we detected the orphan nuclear receptor Nurr1 in these cells. This was followed by a time-dependent up-regulation of the enzymes tyrosine hydroxylase and aldehyde dehydrogenase 2 characteristic of mature dopaminergic neurones. Our in vitro model of dopaminergic cell differentiation corroborates recent in vivo observations in the developing rodent brain.

Citing Articles

The proteome of the differentiating mesencephalic progenitor cell line CSM14.1 in vitro.

Weiss B, Haas S, Lessner G, Mikkat S, Kreutzer M, Glocker M Biomed Res Int. 2014; 2014:351821.

PMID: 24592386 PMC: 3925624. DOI: 10.1155/2014/351821.

Immortalization of neuronal progenitors using SV40 large T antigen and differentiation towards dopaminergic neurons.

Alwin Prem Anand A, Sankar S, Kokila Vani V J Cell Mol Med. 2012; 16(11):2592-610.

PMID: 22863662 PMC: 4118228. DOI: 10.1111/j.1582-4934.2012.01607.x.

Expression patterns of Nurr1 in rat retina development.

Li Y, Qi Q, Cong B, Shi W, Liu X, Zhang G J Mol Histol. 2012; 43(6):633-9.

PMID: 22714110 DOI: 10.1007/s10735-012-9433-z.

Chemical biology investigation of cell death pathways activated by endoplasmic reticulum stress reveals cytoprotective modulators of ASK1.

Kim I, Shu C, Xu W, Shiau C, Grant D, Vasile S J Biol Chem. 2008; 284(3):1593-603.

PMID: 19004820 PMC: 2615512. DOI: 10.1074/jbc.M807308200.

Orthotopic transplantation of immortalized mesencephalic progenitors (CSM14.1 cells) into the substantia nigra of hemiparkinsonian rats induces neuronal differentiation and motoric improvement.

Haas S, Petrov S, Kronenberg G, Schmitt O, Wree A J Anat. 2007; 212(1):19-30.

PMID: 18036147 PMC: 2423384. DOI: 10.1111/j.1469-7580.2007.00834.x.

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