Cell Therapy Manufacturing and Quality Control: Current Process and Regulatory Challenges

Abstract

Mesenchymal stromal cells (MSC) are multipotent stem cells that have been isolated from multiple tissue sources and are currently being used to demonstrate their therapeutic efficacy against various clinical indications. Dramatic increase in the use of MSCs for tissue engineering applications and in regenerative medicine in past two decades raises an increasing demand for cGMP (current Good Manufacturing Practice) based large-scale manufacturing process of MSCs and characterization of these cells. The challenge is to assure the safety and high-quality of cells that will ultimately be therapeutically effective. GMP compliance processing such as cell culture, expansion and cryopreservation is mandatory for making the cell therapy effective. MSCs from various tissue sources should be cultured for scale-up according to regulatory compliance to optimize culture conditions and to ensure the safety of these manufactured cell populations. This review describes cGMP compliances and manufacturing process of bone marrow derived MSCs; specifically in the context of establishing the process flow and in-process controls for the manufacturing process. Importantly, this review highlights the current manufacturing challenges and opportunities for process improvisation and its relevance's for MSCs therapeutics potential.

Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

“…No significant difference was found in hMSC growth in either the FC40 fresh /DMEM or FC40 recycled /DMEM systems with possible cost implications at larger scales. However, the successful development of hMSC manufacturing system is deemed successful if cell quality and identity remain intact . As such, it was imperative to determine if hMSC identity and quality were affected by the perfluorocarbon recycling step.…”

“…However, the successful development of hMSC manufacturing system is deemed successful if cell quality and identity remain intact. 5,48 As such, it was imperative to determine if hMSC identity and quality were affected by the perfluorocarbon recycling step. Figure 8(A) and (B) show the multiparameter flow cytometry analysis of harvested M2 cells after 6 days culture in the two-phase systems when using either fresh or recycled perfluorocarbon, demonstrating the co-expression and lack of expression of the cell surface markers panel recommended by the ISCT.…”

Anti-apoptotic Effects of Human Wharton's Jelly-derived Mesenchymal Stem Cells on Skeletal Muscle Cells Mediated via Secretion of XCL1

“…[30][31][32] Therefore, key secretory proteins of human MSCs are also used directly as novel treatments for patients and could be potent biomarkers for selection of superior human MSCs during large-scale cultivation of clinical grade human MSC in Good Manufacturing Practice facilities. 33 Based on these findings, we aimed to determine if either human MSCs or a selection of key secreted proteins may be applicable as novel treatments. There are many proposed causes of muscle disorders, such as genetic mutation, damage by exercise, aging, mitochondrial defect, and inflammation [6][7][8] and in all the cases, myoblast death or abnormality of myotubes appears to as the final form of muscle disease.…”

TuningForkhead Box D3overexpression to promote specific osteogenic differentiation of human embryonic stem cells while reducing pluripotency in a three‐dimensional culture system

“…Current strategies for guided hESC osteogenic differentiation are limited by (a) the often long and complicated protocols that make them challenging and expensive for clinical implementation (Kim et al, ; Kurosawa, ) and/or (b) the lack of controllable differentiation that increases the risk of heterogeneous and ectopic tissue formation (Heng et al, ; Heslop et al, ) and thus poses challenging safety hurdles (Hourd, Chandra, Medcalf, & Williams, ; Kolkundkar, ). To address these limitations, our long‐term goal is to manipulate the fate of hESCs through overexpression of FOXD3 , a critical regulator of embryonic cell fate, coupled with 3D culture under osteoinductive conditions.…”