Microcarrier expansion of c-MycERTAM -modified human olfactory mucosa cells for neural regeneration.
Microcarrier expansion of c-MycERTAM -modified human olfactory mucosa cells for neural regeneration.
复制标题
c-MycERTAM 修饰的人嗅粘膜细胞的微载体扩增用于神经再生。
DOI：
10.1002/bit.27573
复制
发表时间：
2021
影响因子：
3.8
通讯作者：
Valinhas A
中科院分区：
文献类型：
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作者：
Valinhas A
人嗅粘膜细胞(HOMCs)具有作为脊髓损伤再生治疗的潜力。在我们早期的工作中，我们获得了PA5细胞，这是一个保留了原代人类OMCs功能属性的多克隆群体。微载体悬浮培养是一种替代平面二维培养的方法，可以大量生产满足临床发展需要的细胞。本研究旨在筛选10种微载体对PA5 hOMCs产量和表型的影响。在孔板中进行的研究导致塑料上的细胞产量比塑料+微载体高2.9倍，神经标记物β-III微管蛋白和巢蛋白在两种情况下都上调。微载体悬浮培养7天后，塑料和塑料+细胞的浓度分别为1.4 × 10 5细胞/ml和4.9 × 10 4细胞/ml。与塑料相比，生长在塑料Plus上的PA5 hOMCs的p75NTR转录本显著上调。此外，生长在Platform Plus上的PA5 hOMCs与神经细胞系(NG108-15)共培养后，轴突生长增加。这项研究表明，利用微载体悬浮培养成功地扩增了PA5细胞，并揭示了微载体对细胞扩张和功能属性的竞争效应，表明设计可扩展的生物过程不应该仅仅由细胞产量驱动。
Human olfactory mucosa cells (hOMCs) have potential as a regenerative therapy for spinal cord injury. In our earlier work, we derived PA5 cells, a polyclonal population that retains functional attributes of primary human OMCs. Microcarrier suspension culture is an alternative to planar two‐dimensinal culture to produce cells in quantities that can meet the needs of clinical development. This study aimed to screen the effects of 10 microcarriers on PA5 hOMCs yield and phenotype. Studies performed in well plates led to a 2.9‐fold higher cell yield on plastic compared to plastic plus microcarriers with upregulation of neural markers β‐III tubulin and nestin for both conditions. Microcarrier suspension culture resulted in concentrations of 1.4 × 105cells/ml and 4.9 × 104cells/ml for plastic and plastic plus, respectively, after 7 days. p75NTRtranscript was significantly upregulated for PA5 hOMCs grown on Plastic Plus compared to Plastic. Furthermore, coculture of PA5 hOMCs grown on Plastic Plus with a neuronal cell line (NG108‐15) led to increased neurite outgrowth. This study shows successful expansion of PA5 cells using suspension culture on microcarriers, and it reveals competing effects of microcarriers on cell expansion versus functional attributes, showing that designing scalable bioprocesses should not only be driven by cell yields.