Abstract
The secretome-mediated responses over cellular physiology are well documented. Stem cells have been ruling the field of secretomics and its role in regenerative medicine since the past few years. However, the mechanistic aspects of secretome-mediated responses and the role of other cells in this area remain somewhat elusive. Here, we investigate the effects of secretome-enriched conditioned medium (CM) of neuronally differentiated PC12 cells on the neuronal differentiation of human mesenchymal stem cells (hMSCs). The exposure to CM at a ratio of 1:1 (CM: conditioned medium of PC12 cells) led to neuronal induction in hMSCs. This neuronal induction was compared with a parallel group of cells exposed to nerve growth factor (NGF). There was a marked increase in neurite length and expression of neuronal markers (β-III tubulin, neurofilament-M (NF-M), synaptophysin, NeuN in exposed hMSCs). Experimental group co-exposed to NGF and CM showed an additive response via MAPK signaling and directed the cells particularly towards cholinergic lineage. The ability of CM to enhance the neuronal properties of stem cells could aid in their rapid differentiation into neuronal subtypes in case of stem cell transplantation for neuronal injuries, thus broadening the scope of non-stem cell-based applications in the area of secretomics.
Similar content being viewed by others
References
Tjalsma H, Bolhuis A, Jongbloed JD, Bron S, van Dijl JM (2000) Signal peptide-dependent protein transport in Bacillus subtilis: a genome-based survey of the secretome. Microbiol Mol Biol Rev 64(3):515–547
Karagiannis GS, Pavlou MP, Diamandis EP (2010) Cancer secretomics reveal pathophysiological pathways in cancer molecular oncology. Mol Oncol 4(6):496–510
Garg G, Ranganathan S (2011) In silico secretome analysis approach for next generation sequencing transcriptomic data. BMC Genomics 12(3):S14
Barderas R, Mendes M, Torres S, Bartolomé RA, López-Lucendo M, Villar-Vázquez R, Peláez-García A, Fuente E et al (2013) In-depth characterization of the secretome of colorectal cancer metastatic cells identifies key proteins in cell adhesion, migration, and invasion. Mol Cell Proteomics 12(6):1602–1620
Ditgen D, Anandarajah EM, Meissner KA, Brattig N, Wrenger C, Liebau E (2014) Harnessing the helminth secretome for therapeutic immunomodulators. BioMed Research International 2014, Article ID 964350
Kawahara R, Lima RN, Domingues RR, Pauletti BA, Meirelles GV, Assis M, Figueira ACM, Leme AFP (2014) Deciphering the role of the ADAM17-dependent secretome in cell signaling. J Proteome Res 13(4):2080–2093
Jha MK, Seo M, Kim J-H, Kim B-G, Cho J-Y, Suk K (2013) The secretome signature of reactive glial cells and its pathological implications. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics 1834(11):2418–2428
Cantinieaux D, Quertainmont R, Blacher S, Rossi L, Wanet T, Noël A, Brook G, Schoenen J et al (2013) Conditioned medium from bone marrow-derived mesenchymal stem cells improves recovery after spinal cord injury in rats: an original strategy to avoid cell transplantation. PLoS One 8(8):e69515
Dowling P, Clynes M (2011) Conditioned media from cell lines: a complementary model to clinical specimens for the discovery of disease-specific biomarkers. Proteomics 11(4):794–804
Teixeira FG, Carvalho MM, Panchalingam KM, Rodrigues AJ, Mendes-Pinheiro B, Anjo S, Manadas B, Behie LA et al (2017) Impact of the secretome of human mesenchymal stem cells on brain structure and animal behavior in a rat model of Parkinson’s disease. Stem Cells Transl Med 6(2):634–646
Ribeiro CA, Fraga JS, Grãos M, Neves NM, Reis RL, Gimble JM, Sousa N, Salgado AJ (2012) The secretome of stem cells isolated from the adipose tissue and Wharton jelly acts differently on central nervous system derived cell populations. Stem Cell Res Ther 3(3):18
Chang C-P, Chio C-C, Cheong C-U, Chao C-M, Cheng B-C, Lin M-T (2013) Hypoxic preconditioning enhances the therapeutic potential of the secretome from cultured human mesenchymal stem cells in experimental traumatic brain injury. Clin Sci 124(3):165–176
Salgado AJ, Sousa JC, Costa BM, Pires AO, Mateus-Pinheiro A, Teixeira F, Pinto L, Sousa N (2015) Mesenchymal stem cells secretome as a modulator of the neurogenic niche: basic insights and therapeutic opportunities. Front Cell Neurosci 9:249
Gemma C, Bachstetter AD (2013) The role of microglia in adult hippocampal neurogenesis. Front Cell Neurosci 7:229
Glavaski-Joksimovic A (2014) The mesenchymal stem cell secretome: implications for treatment of traumatic brain injury. Spine 2(1):1007
Makridakis M, Roubelakis MG, Vlahou A (2013) Stem cells: insights into the secretome. Biochim Biophy Acta (BBA)-Proteins and Proteomics 1834(11):2380–2384
Agrawal M, Kumar V, Singh AK, Kashyap MP, Khanna VK, Siddiqui MA, Pant AB (2012) Trans-resveratrol protects ischemic PC12 cells by inhibiting the hypoxia associated transcription factors and increasing the levels of antioxidant defense enzymes. ACS Chem Neurosci 4(2):285–294
Sadri S, Khazaei M, Ghanbari A, Khazaei MR, Shah P (2014) Neuronal differentiation of PC12 and embryonic stem cells in two-and three-dimensional in vitro culture. Indian J Exp Biol 52(4):305–311
Siddiqui M, Kashyap M, Khanna V, Yadav S, Pant A (2010) NGF induced differentiated PC12 cells as in vitro tool to study 4-hydroxynonenal induced cellular damage. Toxicol in Vitro 24(6):1681–1688
Taupenot L (2007) Analysis of regulated secretion using PC12 cells. Curr Prot in Cell Biol:15–12
Vazquez-Martinez R, Almabouada F, Rabanal Y, Diaz-Ruiz A, Garcia-Navarro S, Malagon MM (2013) The small GTPase Rab18 modulates neuroendocrine secretion by interacting with components of the microtubule-based secretory granule transport machinery. Endocr Abstr 32:P683
Kashyap M, Singh A, Siddiqui M, Kumar V, Tripathi V, Khanna V, Yadav S, Jain S et al (2010) Caspase cascade regulated mitochondria mediated apoptosis in monocrotophos exposed PC12 cells. Chem Res Toxicol 23(11):1663–1672
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63
Kumar V, Tripathi VK, Singh AK, Lohani M, Kuddus M (2013) Trans-resveratrol restores the damages induced by organophosphate pesticide-monocrotophos in neuronal cells. Toxicol Int 20(1):48–55
Jahan S, Kumar D, Kumar A, Rajpurohit CS, Singh S, Srivastava A, Pandey A, Pant A (2017) Neurotrophic factor mediated neuronal differentiation of human cord blood mesenchymal stem cells and their applicability to assess the developmental neurotoxicity. Biochem Biophys Res Commun 482(4):961–967
Lee W-C, Chen Y-Y, Kan D, Chien C-L (2012) A neuronal death model: overexpression of neuronal intermediate filament protein peripherin in PC12 cells. J Biomed Sci 19(1):8
Zhu G, Sun L, Keithley RB, Dovichi NJ (2013) Capillary isoelectric focusing-tandem mass spectrometry and reversed-phase liquid chromatography-tandem mass spectrometry for quantitative proteomic analysis of differentiating PC12 cells by eight-plex isobaric tags for relative and absolute quantification. Anal Chem 85(15):7221–7229
Muñoz-Elías G, Woodbury D, Black IB (2003) Marrow stromal cells, mitosis, and neuronal differentiation: stem cell and precursor functions. Stem Cells 21(4):437–448
Tsai H-L, Deng W-P, Lai W-FT, Chiu W-T, Yang C-B, Tsai Y-H, Hwang S-M, Renshaw PF (2014) Wnts enhance neurotrophin-induced neuronal differentiation in adult bone-marrow-derived mesenchymal stem cells via canonical and noncanonical signaling pathways. PLoS One 9(8):e104937
Pires AO, Neves-Carvalho A, Sousa N, Salgado AJ (2014) The secretome of bone marrow and Wharton jelly derived mesenchymal stem cells induces differentiation and neurite outgrowth in SH-SY5Y cells. Stem Cells Int 2014, Article ID 438352
Hsieh J-Y, Wang H-W, Chang S-J, Liao K-H, Lee I-H, Lin W-S, Wu C-H, Lin W-Y et al (2013) Mesenchymal stem cells from human umbilical cord express preferentially secreted factors related to neuroprotection, neurogenesis, and angiogenesis. PLoS One 8(8):e72604
Yamamoto H, Demura T, Morita M, Banker GA, Tanii T, Nakamura S (2012) Differential neurite outgrowth is required for axon specification by cultured hippocampal neurons. J Neurochem 123(6):904–910
Tzeng H-H, Hsu C-H, Chung T-H, Lee W-C, Lin C-H, Wang W-C, Hsiao C-Y, Leu Y-W et al (2015) Cell signaling and differential protein expression in neuronal differentiation of bone marrow mesenchymal stem cells with hypermethylated Salvador/Warts/Hippo (SWH) pathway genes. PLoS One 10(12):e0145542
Xiong Z, Zhao S, Mao X, Lu X, He G, Yang G, Chen M, Ishaq M et al (2014) Selective neuronal differentiation of neural stem cells induced by nanosecond microplasma agitation. Stem Cell Res 12(2):387–399
Montzka K, Lassonczyk N, Tschöke B, Neuss S, Führmann T, Franzen R, Smeets R, Brook GA et al (2009) Neural differentiation potential of human bone marrow-derived mesenchymal stromal cells: misleading marker gene expression. BMC Neurosci 10(1):16
Faigle R, Song H (2013) Signaling mechanisms regulating adult neural stem cells and neurogenesis. Biochim Biophy Acta-Gen Subj 1830(2):2435–2448
Nguyen L, Rigo J-M, Rocher V, Belachew S, Malgrange B, Rogister B, Leprince P, Moonen G (2001) Neurotransmitters as early signals for central nervous system development. Cell Tissue Res 305(2):187–202
Hyman SE (2005) Neurotransmitters. Curr Biol 15(5):R154–R158
Foster AC, Kemp JA (2006) Glutamate-and GABA-based CNS therapeutics. Curr Opin Pharmacol 6(1):7–17
Hangya B, Ranade SP, Lorenc M, Kepecs A (2015) Central cholinergic neurons are rapidly recruited by reinforcement feedback. Cell 162(5):1155–1168
Hynes M, Porter JA, Chiang C, Chang D, Tessier-Lavigne M, Beachy PA, Rosenthal A (1995) Induction of midbrain dopaminergic neurons by Sonic hedgehog. Neuron 15(1):35–44
Kanunnikova NP (2012) Role of brain glutamic acid metabolism changes in neurodegenerative pathologies. J Biol Earth Sci 2012 2(1):10
Kegeles LS (2016) Brain GABA function and psychosis. Am J Psychiatr 173(5):448–449
Kegeles LS, Abi-Dargham A, Frankle WG, Gil R, Cooper TB, Slifstein M, Hwang D-R, Huang Y et al (2010) Increased synaptic dopamine function in associative regions of the striatum in schizophrenia. Arch Gen Psychiatry 67(3):231–239
Moret C, Briley M (2011) The importance of norepinephrine in depression. Neuropsychiatr Dis Treat 7(Suppl 1):9–13
Szutowicz A, Bielarczyk H, Jankowska-Kulawy A, Pawełczyk T, Ronowska A (2013) Acetyl-CoA the key factor for survival or death of cholinergic neurons in course of neurodegenerative diseases. Neurochem Res 38(8):1523–1542
Liang J, Wu S, Zhao H, Li S-l, Liu Z-x, Wu J, Zhou L (2013) Human umbilical cord mesenchymal stem cells derived from Wharton’s jelly differentiate into cholinergic-like neurons in vitro. Neurosci Lett 532:59–63
Chinta SJ, Andersen JK (2005) Dopaminergic neurons. Int J Biochem Cell Biol 37(5):942–946
Campos-Peña V, Meraz-Ríos MA (2014) Alzheimer disease: the role of Aβ in the glutamatergic system. In: Neurochemistry. InTech. https://doi.org/10.5772/57367
Ranganath SH, Levy O, Inamdar MS, Karp JM (2012) Harnessing the mesenchymal stem cell secretome for the treatment of cardiovascular disease. Cell Stem Cell 10(3):244–258
Huang EJ, Reichardt LF (2003) Trk receptors: roles in neuronal signal transduction. Annu Rev Biochem 72(1):609–642
Lee FS, Chao MV (2001) Activation of Trk neurotrophin receptors in the absence of neurotrophins. Proc Natl Acad Sci 98(6):3555–3560
Lemmon MA, Schlessinger J (2010) Cell signaling by receptor tyrosine kinases. Cell 141(7):1117–1134
Houslay MD, Kolch W (2000) Cell-type specific integration of cross-talk between extracellular signal-regulated kinase and cAMP signaling. Mol Pharmacol 58(4):659–668
Sweatt JD (2001) The neuronal MAP kinase cascade: a biochemical signal integration system subserving synaptic plasticity and memory. J Neurochem 76(1):1–10
Oeztuerk-Winder F, Ventura J-J (2012) The many faces of p38 mitogen-activated protein kinase in progenitor/stem cell differentiation. Biochem J 445(1):1–10
Chen F (2012) JNK-induced apoptosis, compensatory growth, and cancer stem cells. Cancer Res 72(2):379–386
Kumar V, Gupta AK, Shukla RK, Tripathi VK, Jahan S, Pandey A, Srivastava A, Agrawal M et al (2015) Molecular mechanism of switching of TrkA/p75NTR signaling in monocrotophos induced neurotoxicity. Sci Rep 5:14038
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Srivastava, A., Singh, S., Pandey, A. et al. Secretome of Differentiated PC12 Cells Enhances Neuronal Differentiation in Human Mesenchymal Stem Cells Via NGF-Like Mechanism. Mol Neurobiol 55, 8293–8305 (2018). https://doi.org/10.1007/s12035-018-0981-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12035-018-0981-4