Insights in Enzyme Research

Differentiation of mesenchymal stem cells into nucleus pulposus like cells induced by co-culture system and hypoxia

EuroSciCon Congress on Enzymology and Molecular Biology
August 13-14, 2018 Paris ,France

Arjun Sinkemani, Feng Wang, Zhi-Yang Xie, Lu Chen, Cong Zhang and Xiao-Tao Wu

Zhongda Hospital, Southeast University, Nanjing, China

ScientificTracks Abstracts: Insights Enzyme Res

DOI: 10.21767/2573-4466-C1-002

Abstract

Introduction: Intervertebral disc (IVD) is closely related to low back pain, is the major cause of disability worldwide, with more than 84% population experiencing pain in their life time. Biological and cell based therapies are on progress as an optional treatment for IVD degeneration. Many studies have also revealed that mesenchymal stem cells (MSCs) can also be differentiated into nucleus pulposus (NP) like cells phenotype. This study aimed to determine the newly defined healthy NP cells markers; HIF- 1α, HIF-2α, GLUT-1, Shh, Brachyury, Aggrecan, Collagen II, Carbonic anhydrase 3, Carbonic anhydrase 12, CD24, Cytokeratin 8, Cytokeratin 18, and Cytokeratin 19 of Sprague-Dawley rat, whether these markers can be expressed in MSCs under co-culture condition and could be identified the differentiation of MSCs into NP-like cells. Methods: NP cells and bone marrow derived MSCs from Sprague-Dawley rats were cultured under normoxic medium at 21% O2 and 5% CO2 at 37�?�?C and hypoxic medium at 2% O2, 5% CO2, 93% N2 at 37�?�?C and MSCs were co-cultured with NP cells supernatant with the concentration of 50% and 100% for 7 days under both normoxic and hypoxic medium. Differentiation of MSCs and expression of recommended newly defined young healthy NP cells phenotypes were evaluated by quantitative real-time PCR (qPCR), Western blotting and immunofluorescence staining microscopy. The results were determined among the groups using unpaired Student’s t-test. p-values<0.05 considered significant. Results: MSCs co-cultured with the concentration of 50% NPcs supernatant; only collagen II showed the increased expression while with the 100% NPcs supernatant; brachyury, collagen II, Glut-1, KRT18 and KRT19 showed higher expressions under normoxic condition compared to MSC control. Under the hypoxic condition, MSCs co-cultured with 50% NPcs supernatant, HIF- 2α, Glut-1, aggrecan, collagen II, shh, KRT8, KRT19, CA3, CA12 and CD24 showed increased expression compared to MSC control. More importantly, MSCs co-cultured with 100% NPcs supernatant under hypoxic condition, HIF-1α, HIF-2α, Glut-1, aggrecan, collagen II, shh, brachyury, KRT8, KRT19, CA3, CA12 and CD24 showed upregulated increased expressions compared to the MSC control, which showed that NP cells can stimulate MSCs differentiation to NP-like cells with paracrine interaction between MSCs and NPcs under co-culture condition. Conclusion: This study suggested that MSCs

Biography

Arjun sinkeman, has completed his Master’s degree in Orthopaedic Surgery from Southeast University and now pursuing his PhD study in the same university. He has more than 5 scientific publications. He has participated in the 20th Asia Pacific Orthopaedic Association Congress, Turkey- Oral Presentation; the 12th International Congress of Chinese Orthopaedic Association, China- E-Poster Presentation; A O Spine Advanced Symposium, China- Controversial Case Discussion Forum; Orthopaedic Research Society 47th International Musculoskeletal Biology Workshop, USA- Poster Presentation; International Symposium on Life Science & Biological Engineering, Hong Kong- Oral Presentation; International Symposium on Life Science & Biological Engineering, Japan- Oral Presentation and the 10th International Congress of Chinese Orthopaedic Association, China. He holds the active Memberships of A O Spine, Orthopaedic Research Society and North American Spine Society. He is awarded with the Chinese Government Scholarship for outstanding international students; Nanjing Municipal Government International Students Scholarship; 1st Category Southeast University Scholarship and Chinese Government Scholarship.

E-mail: sinkemani@hotmail.com