Microcarrier expansion of c-MycERTAM - modified human olfactory mucosa cells for neural regeneration.,Biotechnology and Bioengineering

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Microcarrier expansion of c-MycERTAM - modified human olfactory mucosa cells for neural regeneration.
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2020-09-21 , DOI: 10.1002/bit.27573
Ana Valinhas ₁ , Gerardo Santiago-Toledo ₁ , Ivan B Wall _{1,

2,

3}

Affiliation

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 × 10⁵ cells/ml and 4.9 × 10⁴ cells/ml for plastic and plastic plus, respectively, after 7 days. p75^NTR transcript 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.

中文翻译：

c-MycERTAM 的微载体扩增 - 用于神经再生的修饰的人类嗅觉粘膜细胞。

人类嗅觉粘膜细胞 (hOMC) 具有作为脊髓损伤再生疗法的潜力。在我们早期的工作中，我们衍生了 PA5 细胞，这是一种保留原代人类 OMC 功能属性的多克隆细胞群。微载体悬浮培养是平面二维培养的替代方案，可生产满足临床开发需要的细胞数量。本研究旨在筛选 10 种微载体对 PA5 hOMCs 产量和表型的影响。与塑料加微载体相比，在孔板中进行的研究使塑料上的细胞产量提高了 2.9 倍，两种条件下神经标志物 β-III 微管蛋白和巢蛋白均上调。微载体悬浮培养导致浓度为 1.4 × 10 ^{5 个}细胞/ml 和 4.9 × 10 ⁴7 天后分别为塑料和塑料加的细胞/毫升。与塑料相比，在塑料 Plus 上生长的 PA5 hOMC 的p75 ^NTR转录物显着上调。此外，在 Plastic Plus 上生长的 PA5 hOMC 与神经元细胞系 (NG108-15) 共培养导致神经突生长增加。该研究表明，在微载体上使用悬浮培养成功扩增了 PA5 细胞，并揭示了微载体对细胞扩增与功能属性的竞争影响，表明设计可扩展的生物过程不应仅由细胞产量驱动。

更新日期：2020-09-21

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