Wojciech Fortuna1, Benita Wiatrak2, Paulina Jawień3, Adriana Kubis-Kubiak4, Ying Li5, Daqing Li5, Paweł Tabakow1. 1. Department of Neurosurgery, Wroclaw Medical University, Wroclaw, Poland. 2. Department of Pharmacology, Wroclaw Medical University, Wroclaw, Poland; benita.wiatrak@umw.edu.pl. 3. Department of Biostructure and Animal Physiology, Wroclaw University of Environmental and Life Sciences, Wroclaw, Poland. 4. Department of Toxicology, Faculty of Pharmacy, Wroclaw Medical University, Wroclaw, Poland. 5. Spinal Repair Unit, Department of Brain Repair and Rehabilitation, UCL Institute of Neurology, London, U.K.
Abstract
BACKGROUND/AIM: The regeneration of a completely damaged spinal cord is still a challenge in modern medicine. A promising treatment method is autologous transplantation of olfactory ensheathing cells (OECs). This study aimed primarily to test methods of culturing OECs with the use of materials and reagents that are certified for pharmaceutical use in the production of an advanced cell therapy product intended for humans. MATERIALS AND METHODS: The culture of OECs was performed using various modifications of the surface of the culture vessels (with fibronectin and poly-D-lysine). The number of cells was assessed after immunofluorescence staining using anti-fibronectin and anti-p75 NGF receptor antibodies. The study compared, in terms of surgical manipulations, scaffolds with OECs prepared based on 3 types of collagen: Acid Solubilized Telo Collagen and Pepsin Solubilized Atelocollagen, and the popular Corning collagen. RESULTS: We have shown that when suspending OECs in collagen gel, it is much better to use acid-solubilized collagen (ASC) than pepsin-solubilized collagen (PSC) because the 3D collagen scaffold from ASC provides much easier handling of the product during a surgical procedure. We also found that the OEC cultures should be grown on the surface modified with fibronectin. Furthermore, we have also shown that the optimal concentration of fetal bovine serum (FBS) for culturing these cells should be around 10%. CONCLUSION: The culture of OECs based on reagents intended for human use can be successfully carried out, obtaining sufficient OECs content in the heterogeneous cell culture to produce a functional advanced therapy medicinal product.
BACKGROUND/AIM: The regeneration of a completely damaged spinal cord is still a challenge in modern medicine. A promising treatment method is autologous transplantation of olfactory ensheathing cells (OECs). This study aimed primarily to test methods of culturing OECs with the use of materials and reagents that are certified for pharmaceutical use in the production of an advanced cell therapy product intended for humans. MATERIALS AND METHODS: The culture of OECs was performed using various modifications of the surface of the culture vessels (with fibronectin and poly-D-lysine). The number of cells was assessed after immunofluorescence staining using anti-fibronectin and anti-p75 NGF receptor antibodies. The study compared, in terms of surgical manipulations, scaffolds with OECs prepared based on 3 types of collagen: Acid Solubilized Telo Collagen and Pepsin Solubilized Atelocollagen, and the popular Corning collagen. RESULTS: We have shown that when suspending OECs in collagen gel, it is much better to use acid-solubilized collagen (ASC) than pepsin-solubilized collagen (PSC) because the 3D collagen scaffold from ASC provides much easier handling of the product during a surgical procedure. We also found that the OEC cultures should be grown on the surface modified with fibronectin. Furthermore, we have also shown that the optimal concentration of fetal bovine serum (FBS) for culturing these cells should be around 10%. CONCLUSION: The culture of OECs based on reagents intended for human use can be successfully carried out, obtaining sufficient OECs content in the heterogeneous cell culture to produce a functional advanced therapy medicinal product.
Authors: Jeff Biernaskie; Joseph S Sparling; Jie Liu; Casey P Shannon; Jason R Plemel; Yuanyun Xie; Freda D Miller; Wolfram Tetzlaff Journal: J Neurosci Date: 2007-09-05 Impact factor: 6.167
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Authors: R Yao; M Murtaza; J Tello Velasquez; M Todorovic; A Rayfield; J Ekberg; M Barton; J St John Journal: Cell Transplant Date: 2018-06-08 Impact factor: 4.064
Authors: A Mackay-Sim; F Féron; J Cochrane; L Bassingthwaighte; C Bayliss; W Davies; P Fronek; C Gray; G Kerr; P Licina; A Nowitzke; C Perry; P A S Silburn; S Urquhart; T Geraghty Journal: Brain Date: 2008-08-08 Impact factor: 13.501