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Neurogenic transdifferentiation of human adipose-derived stem cells? A critical protocol reevaluation with special emphasis on cell proliferation and cell cycle alterations

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Abstract

Adipose-derived stem cells (ASCs) are reported to display multilineage differentiation potential, including neuroectodermal pathways. The aim of the present study was to critically re-evaluate the potential neurogenic (trans-)differentiation capacity of ASCs using a neurogenic induction protocol based on the combination of isobutylmethylxanthine (IBMX), indomethacin and insulin. ASCs isolated from lipo-aspirate samples of five healthy female donors were characterized and potential neurogenic (trans-)differentiation was assessed by means of immunohistochemistry and gene expression analyses. Cell proliferation and cell cycle alterations were studied, and the expression of CREB/ATF transcription factors was analyzed. ASCs expressed CD59, CD90 and CD105, and were tested negative for CD34 and CD45. Under neurogenic induction, ASCs adopted a characteristic morphology comparable to neur(on)al progenitors and expressed musashi1, β-III-tubulin and nestin. Gene expression analyses revealed an increased expression of β-III-tubulin, GFAP, vimentin and BDNF, as well as SOX4 in induced ASCs. Cell proliferation was significantly reduced under neurogenic induction; cell cycle analyses showed a G2-cell cycle arrest accompanied by differential expression of key regulators of cell cycle progression. Differential expression of CREB/ATF transcription factors could be observed on neurogenic induction, pointing to a decisive role of the cAMP-CREB/ATF system. Our findings may point to a potential neurogenic (trans-)differentiation of ASCs into early neur(on)al progenitors, but do not present definite evidence for it. Especially, the adoption of a neural progenitor cell-like morphology must not automatically be misinterpreted as a specific characteristic of a respective (trans-)differentiation process, as this may as well be caused by alterations of cell cycle progression.

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Abbreviations

ASC:

Adipose-derived stem cell(s)

ATF:

Activating transcription factor

bCM:

Basic culture medium

BMSC:

Bone marrow-derived mesenchymal stromal (stem) cells

cAMP:

Cyclic adenosine monophosphate

CREB:

cAMP response element-binding protein

CREM:

cAMP response element modulator

ESC:

Embryonic stem cells

IBMX:

Isobutylmethylxanthine

NID:

Neurogenic induction medium

SVF:

Stromal vascular fraction

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Acknowledgments

The authors especially would like to thank R. Gehrcke and T. Jansen for their support.

Conflict of interest

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Anatomy II: Experimental Morphology, Center for Experimental Medicine, University Hospital Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany

    Kai Michael Kompisch, Reinhard Müller & Udo Schumacher

  2. Clinic for Stem Cell Transplantation, Center of Oncology, University Hospital Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany

    Claudia Lange

  3. Institute for Cellular and Molecular Pathology, Center for Pathology and Forensic and Genetic Medicine, Medizinische Hochschule Hannover, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Doris Steinemann, Britta Skawran & Brigitte Schlegelberger

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  1. Kai Michael Kompisch
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  2. Claudia Lange
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Correspondence to Kai Michael Kompisch.

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Kompisch, K.M., Lange, C., Steinemann, D. et al. Neurogenic transdifferentiation of human adipose-derived stem cells? A critical protocol reevaluation with special emphasis on cell proliferation and cell cycle alterations. Histochem Cell Biol 134, 453–468 (2010). https://doi.org/10.1007/s00418-010-0740-8

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