Bioprocessing strategies to enhance the challenging isolation of neuro-regenerative cells from olfactory mucosa.
Bioprocessing strategies to enhance the challenging isolation of neuro-regenerative cells from olfactory mucosa.
复制标题
DOI：
10.1038/s41598-018-32748-w
复制
发表时间：
2018-09-27
影响因子：
4.6
通讯作者：
Wall I
中科院分区：
文献类型：
--
作者：
Georgiou M;Reis JND;Wood R;Esteban PP;Roberton V;Mason C;Li D;Li Y;Choi D;Wall I
嗅鞘细胞（OECs）是一种有前途的潜在细胞治疗，以帮助再生。然而，在分离和定性方面存在重大挑战。在目前的研究中，我们已经探索了方法，以提高从大鼠粘膜中OEC标记物p75 NTR表达细胞的回收率。加入24小时的差异粘附步骤后，PDL和层粘连蛋白基质上p75 NTR的表达分别显著增加至73 ± 5%和46 ± 18%。此外，神经营养因子NT-3的引入和血清浓度降低至2%FBS导致OEC富集，p75 NTR接近100%（PDL和层粘连蛋白上分别为100 ± 0%和98 ± 2%），候选成纤维细胞标记物Thy1.1降低至零。在生理相关的氧张力（2-8%）下培养OEC对p75 NTR表达和总体细胞存活具有负面影响。关于细胞效能，OEC与NG 108 -15神经元的共培养导致在大气氧下更多的神经元生长和潜在的迁移。此外，OEC的表现类似于雪旺细胞系阳性对照。总之，这项工作确定了关键的生物加工基础，这些基础将支持未来基于OEC的细胞疗法在脊髓损伤修复中的潜在应用。然而，要创建优化的隔离方法，还有很多工作要做。
Olfactory ensheathing cells (OECs) are a promising potential cell therapy to aid regeneration. However, there are significant challenges in isolating and characterizing them. In the current study, we have explored methods to enhance the recovery of cells expressing OEC marker p75NTR from rat mucosa. With the addition of a 24-hour differential adhesion step, the expression of p75NTR was significantly increased to 73 ± 5% and 46 ± 18% on PDL and laminin matrices respectively. Additionally, the introduction of neurotrophic factor NT-3 and the decrease in serum concentration to 2% FBS resulted in enrichment of OECs, with p75NTR at nearly 100% (100 ± 0% and 98 ± 2% on PDL and laminin respectively), and candidate fibroblast marker Thy1.1 decreased to zero. Culturing OECs at physiologically relevant oxygen tension (2–8%) had a negative impact on p75NTR expression and overall cell survival. Regarding cell potency, co-culture of OECs with NG108-15 neurons resulted in more neuronal growth and potential migration at atmospheric oxygen. Moreover, OECs behaved similarly to a Schwann cell line positive control. In conclusion, this work identified key bioprocessing fundamentals that will underpin future development of OEC-based cell therapies for potential use in spinal cord injury repair. However, there is still much work to do to create optimized isolation methods.
登录
查看更多内容
影响因子：
6.2
作者：
DOUCETTE, R
通讯作者：
DOUCETTE, R
自体 OM-MSC 和 OM-OEC 联合移植：治疗脊髓损伤的新方法
影响因子：
4.1
作者：
Ge, Lite;Liu, Kai;Lu, Ming
通讯作者：
Lu, Ming
影响因子：
3.3
作者：
GRAZIADEI, PPC;LEVINE, RR;MONTIGRAZIADEI, GA
通讯作者：
MONTIGRAZIADEI, GA
影响因子：
2.8
作者：
GudinoCabrera, G;NietoSampedro, M
通讯作者：
NietoSampedro, M
影响因子：
8
作者：
Chung, RS;Woodhouse, A;Chuah, MI
通讯作者：
Chuah, MI