The effect of Noggin supplementation in Matrigel nanofiber-based cell culture system for derivation of neural-like cells from human endometrial-derived stromal cells Shima Tavakol, 1* Sayed Mostafa Modarres Mousavi, 2,3* Mohammad Masummi, 4,5 Amir Amani, 1 Seyed Mahdi Rezayat, 1,6,7 Jafar Ai 8,9 1 Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran 2 Shefa Neuroscience Research Center, Khatam al-Anbia Hospital, Tehran, Iran 3 Shiraz University, Shiraz, Iran 4 Induced Pluripotent Stem Cell Biotechnology Team, Department of Nanobiomaterials and Tissue Engineering, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran 5 Stem Cells Biology Department, Stem Cell Technology Research Center, Tehran, Iran 6 Department of Toxicology and Pharmacology, School of Pharmacy, Pharmaceutical Sciences Branch, Islamic Azad University (IAUPS), Tehran, Iran 7 Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran 8 Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran 9 Brain and Spinal Injury Research Center, Imam Hospital, Tehran University of Medical Sciences, Tehran, Iran Received 9 June 2013; revised 6 November 2013; accepted 31 December 2013 Published online 00 Month 2014 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.a.35079 Abstract: A very important obstacle in axonal regeneration after spinal cord injury is astroglial scaring. Noggin as bone morphogenic protein inhibitor plays a critical role in decreas- ing GFAP 1 cells and reducing the number of astrocytes in the site of injury. Human endometrial-derived stromal cells (hEnSCs) were isolated and cultured in two different neural inductive mediums consisting of neural progenitor mainte- nance medium (NPMM)/BDNF or NPMM/BDNF/Noggin in Matrigel 3D cell culture. Neural expression markers were investigated at the mRNA and protein level by real-time PCR and immunocytochemistry, respectively. The results showed that Noggin supplementation was able to increase the expression of Nestin, Tuj-1, and NF, whereas the expressions of GFAP, Bcl2, and Olig2 were decreased. In addition, DAPI staining demonstrated that lighter blue chromatin agreed with our observation of lower level of Bcl2 expression in the Noggin protocol in which over-expression of Bcl2 gene did not induce higher neurogenesis in poor Noggin medium. Our findings clearly demonstrated the neural differentiation potential of hEnSC in Matrigel and also Noggin supplementation was able to inhibit astrocyte formation. V C 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 00A:000–000, 2014. Key Words: Matrigel, human endometrial-derived stromal cells, neural differentiation, Noggin, GFAP How to cite this article: Tavakol S, Mousavi SMM, Masummi M, Amani A, Rezayat SM, Ai J. 2014. The effect of Noggin sup- plementation in Matrigel nanoﬁber-based cell culture system for derivation of neural-like cells from human endometrial- derived stromal cells. J Biomed Mater Res Part A 2014:00A:000–000. INTRODUCTION Spinal cord injury is deﬁned as a loss of axon integrity that could result in persistent deﬁcit. One of the inhibitors of axonal regeneration is astroglial scarring. 1 Glial ﬁbrillary acidic protein (GFAP), an intermediate ﬁlament protein and a marker of astrocyte, is expressed in reactive astrocyte in lesions leading to astroglial scarring. Inﬂammation and low cell viability in the CNS inhibit neural differentiation. Low cell viability is one of the major problems for a successful cell replacement therapy. To over- come this obstacle, use of Matrigel as an extracellular matrix has been suggested. There are several methods of nanoﬁbers synthesis including self-assembling. Matrigel con- sists of laminin as the major component heparin sulfate, col- lagen IV, proteoglycans, entactin, and nidogen and forms nanoﬁber at temperatures above 20  C. 2,3 Uemura et al. demonstrated that Matrigel can indirectly increase neural differentiation by reducing local inﬂammation, decrease of CD45-positive cells and protection of neural cells from cyto- toxic inﬂammatory cytokine exposure, such as TNF-a, IL-6, *These authors contributed equally to this work. Correspondence to: J. Ai; e-mail: Jafar_ai@tums.ir.com or S.M. Rezayat; e-mail: rezayat@sina.tums.ac.ir V C 2014 WILEY PERIODICALS, INC. 1