Serum-free bioprocessing of adult human and rodent skin-derived Schwann cells: implications for cell therapy in nervous system injury

Abstract

Peripheral nerve injury affects 2.8% of trauma patients with severe cases often resulting in long-lived permanent disability, despite nerve repair surgery. Autologous Schwann cell (SC) therapy currently provides an exciting avenue for improved outcomes for these patients, particularly with the possibility to derive SCs from easily-accessible adult skin. However, due to current challenges regarding the efficient expansion of these cells, further optimization is required before they can be seriously considered for clinical application. Here, a microcarrier-based bioreactor system is proposed as a means to scale-up large numbers of adult skin-derived SCs for transplantation into the injured nerve. Bioprocessing parameters that allow for the expansion of adult rodent SCs have been identified, whilst maintaining similar rates of proliferation (as compared to static-grown SCs), expression of SC markers, and, importantly, their capacity to myelinate axons following transplant into the injured sciatic nerve. The same bioprocessing parameters can be applied to SCs derived from adult human skin, and like rodent cells, they sustain their proliferative potential and expression of SC markers. Taken together, this dataset demonstrates the basis for a scalable bioprocess for the production of SCs, an important step towards clinical use of these cells as an adjunct therapy for nerve repair. Copyright © 2017 John Wiley & Sons, Ltd.

Purification and Characterization of Schwann Cells from Adult Human Skin and Nerve

“…Interestingly, this was in stark contrast to transplanted adult rodent SCs where ∼80% of transplanted cells exhibited mature features of myelin following identical processing as human cells, and transplanted and assessed in the same model (Kumar et al, 2016; Mirfeizi et al, 2017). Such findings suggest that species differences between donor cells versus recipient may play a role in low myelination efficiency of human cells.…”

Factors Within the Endoneurial Microenvironment Act to Suppress Tumorigenesis of MPNST

“…Sciatic nerves were collected from three adult Sprague Dawley rats, then Schwann cells were isolated using a modified protocol from previously described methods ( Mirfeizi et al, 2017 ). Briefly, nerves were dissected and minced, and subsequently incubated in collagenase Type IV (Worthington) at 37°C for 40 min.…”

“…To track cells following in vivo transplantation, cells were transduced with a GFP-expressing lenti-viral vector as previously described ( Mirfeizi et al, 2017 ). All lentivirus work was undertaken in a biohazard level 2+ laboratory using previously established protocols.…”

GMP Production and Scale-Up of Adherent Neural Stem Cells with a Quantum Cell Expansion System

“…Current scale-up options for adherent cells include multiplate-based technologies, microcarrier systems, and packed-bed bioreactors. 1 , 2 , 3 , 4 The most common method used for adherent cell expansion is multilayer cell factories. However, this approach has several drawbacks, including the amount of time required for expansion, need for multiple passages, use of an open system, and a limited final cell yield due to availability of incubator space and specialized personnel.…”