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Int. J. Mol. Sci. 2015, 16(1), 420-438; doi:10.3390/ijms16010420

Effects of Different Sera Conditions on Olfactory Ensheathing Cells in Vitro

1
Second Department of Orthopedics, Second Affiliated Hospital of Xi’an Jiaotong University, 157 West Five Road, Xincheng District, Xi’an 710004, Shaanxi, China
2
Department of Endocrinology, First Affiliated Hospital of Xi’an Jiaotong University, Yanta West Road, No. 277, Xi’an 710061, China
3
Department of Plastic Surgery, Second Affiliated Hospital of Xi’an Jiaotong University, West Five Road, No. 157, Xi’an 710004, China
4
Department of Orthopedics, Xi’an Honghui hospital of Xi’an Jiaotong University, Nanguo Road, No. 76, Xi’an 710054, China
*
Author to whom correspondence should be addressed.
Academic Editor: Katalin Prokai-Tatrai
Received: 24 June 2014 / Accepted: 17 December 2014 / Published: 26 December 2014
(This article belongs to the Special Issue Neuroprotective Strategies 2014)
View Full-Text   |   Download PDF [4155 KB, uploaded 26 December 2014]   |  

Abstract

Transplantation of olfactory ensheathing cells (OEC) is a promising therapy in spinal cord injury (SCI) treatment. However, the therapeutic efficacy of this method is unstable due to unknown reasons. Considering the alterations in the culture environment that occur during OEC preparation for transplantation, we hypothesize that these changes may cause variations in the curative effects of this method. In this study, we compared OEC cultured in medium containing different types and concentrations of serum. After purification and passage, the OEC were cultured for 7 days in different media containing 5%, 10%, 15% or 20% fetal bovine serum (FBS) or rat serum (RS), or the cells were cultured in FBS-containing medium first, followed by medium containing RS. In another group, the OEC were first cultured in 10% FBS for 3 days and then cultured with rat spinal cord explants with 10% RS for another 4 days. An MTT assay and P75 neurotrophin receptor immunofluorescence staining were used to examine cell viability and OEC numbers, respectively. The concentration of neurotrophin-3 (NT-3), which is secreted by OEC into the culture supernatant, was detected using the enzyme-linked immunosorbent assay (ELISA). RT-PCR was applied to investigate the NT-3 gene expression in OEC according to different groups. Compared with FBS, RS reduced OEC proliferation in relation to OEC counts (χ2 = 166.279, df = 1, p < 0.01), the optical density (OD) value in the MTT assay (χ2 = 34.730, df = 1, p < 0.01), and NT-3 concentration in the supernatant (χ2 = 242.997, df = 1, p < 0.01). OEC cultured with spinal cord explants secreted less NT-3 than OEC cultured alone (F = 9.611, df = 5.139, p < 0.01). Meanwhile, the order of application of different sera was not influential. There was statistically significant difference in NT-3 gene expression among different groups when the serum concentration was 15% (χ2 = 64.347, df = 1, p < 0.01). In conclusion, different serum conditions may be responsible for the variations in OEC proliferation and function. View Full-Text
Keywords: rat serum; fetal bovine serum; olfactory ensheathing cells; concentration; culture rat serum; fetal bovine serum; olfactory ensheathing cells; concentration; culture
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Lu, M.; Dong, J.; Lu, T.; Lv, H.; Yang, P.; Cheng, Z.; Li, J.; Liang, B.; Xu, J.; Li, H.; He, X. Effects of Different Sera Conditions on Olfactory Ensheathing Cells in Vitro. Int. J. Mol. Sci. 2015, 16, 420-438.

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