Microcarrier expansion of c‐MycERTAM‐modified human olfactory mucosa cells for neural regeneration. Issue 1 (7th October 2020)

• Record Type: Journal Article
• Authors: Valinhas, Ana; Santiago‐Toledo, Gerardo; Wall, Ivan B.
• Is Part Of: Biotechnology and bioengineering. Volume 118:Issue 1(2021)
• Page Start: 329
• Page End: 344
• Publication Date: 2020-10-07
• Subjects: allogeneic cell therapy -- human olfactory mucosa cells -- microcarrier -- spinal cord injury -- suspension culture; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.27573
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: 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 × 10 5 cells/ml and 4.9 × 10 4 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. Abstract : – Expanding olfactory ensheathing cells (OECs) for regenerative medicine applications is challenging. – The current study examined the expansion in a stirred system of OECs containing a c‐mycER transgene that can be driven by 4‐OHT. – By selecting the micro carriers that promote best expansion and expression of identity markers it is envisaged that a stable product will be achieved.