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The conditioned medium from a stable human GDF3-expressing CHO cell line, induces the differentiation of PC12 cells

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Abstract

Members of the transforming growth factor-β (TGF-β) superfamily have significant roles in the regulation of a wide variety of physiological processes. In our present work, phylogenetic tree analysis showed that human GDF3 (Growth and differentiation factor 3) and human GDF1 formed a subgroup of closely related molecules. Through quantitative real-time PCR analysis in different human tissues, GDF1 and GDF3 expression level had a big difference in brain. GDF3 could activate downstream signaling through associating with ALK7 (Activin receptor-like kinase 7) in a Cripto-dependent fashion. A CHO cell line stably transfected with the encoding sequence of GDF3, named CHO-GDF3, was established. Western blotting analysis demonstrated that GDF3 protein could be secreted into the medium from CHO cells and immunofluorescence experiment showed that GDF3 was mainly distributed in cytoplasm of the stable cell line, the primary hippocampal neurons, and brain tissues. Furthermore, the conditioned medium from CHO-GDF3 could reduce PC12 cell growth and induce cell differentiation. All these findings bring new insights into the functional study of GDF3.

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Abbreviations

TGF-β superfamily:

Transforming growth factor-β superfamily

GDF3:

Growth and differentiation factor 3

ALK7:

Activin receptor like-kinase 7

BMP:

Bone morphogenetic protein

β2-MG:

β2-macroglobulin

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Acknowledgments

This study was supported by the National 973 program of China (2004CB518605), the National 863 project of China (2006AA020501), the National Key Sci-Tech Special Project of China (2008ZX10002-020), the Project of the Shanghai Municipal Science and Technology Commission (03dz14086), and the National Natural Science foundation of China (30024001, 30771188).

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Authors and Affiliations

  1. State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai, 200433, People’s Republic of China

    Qiang Li, **anghua Liu, Yanhua Wu, Jian An, Saiyin Hexige, Yichen Ling, Mingjun Zhang, **anmei Yang & Long Yu

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  1. Qiang Li
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  2. **anghua Liu
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Correspondence to Qiang Li or Long Yu.

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Li, Q., Liu, X., Wu, Y. et al. The conditioned medium from a stable human GDF3-expressing CHO cell line, induces the differentiation of PC12 cells. Mol Cell Biochem 359, 115–123 (2012). https://doi.org/10.1007/s11010-011-1005-0

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  • DOI: https://doi.org/10.1007/s11010-011-1005-0

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