Quality Control Assays for Clinical-Grade Human Mesenchymal Stromal Cells: Validation Strategy

Abstract

The present chapter focuses on the validation of the following analytical methods for the control of mesenchymal stromal cells (MSC) for cell therapy clinical trials: Microbiological control for cellular product Endotoxin assay Mycoplasma assay Cell count and viability Immunophenotype Clonogenic potential (CFU-F assay) In our lab, these methods are in use for product release, process control or control of the biological starting materials. They are described in detail in the accompanying Chapter 19.For each method, validation goals and strategy are presented, and a detailed experimental scheme is proposed.

Tips and Tricks for Validation of Quality Control Analytical Methods in Good Manufacturing Practice Mesenchymal Stromal Cell Production

“…Nevertheless, different automated methods are available on the market that allow to obtain operator-independent cell counting that in a GMP settings becomes extremely relevant to rely on a univocal accurate test. Gunetti et al [ 26 ] explain in detail the validation of a disposable device in comparison with the Burker chamber, while Radrizzani et al [ 5 ] performed validation of a flow cytometry method.…”

“…In particular, according to International Conference on Harmonization Q2 (ICH Q2 R1) Guidelines [ 4 ], validation of each analytical method is required with the purpose to demonstrate that the procedures and the test adopted from the quality control laboratory are suitable for the intended use, so they are appropriate to give results in terms of quality attributes, as described above. A validation activity is generally composed of four steps: (1) qualification of personnel and equipment used as prerequisite for all the operations; (2) description of the validation strategy in written and approved validation protocols; (3) performance of the validation experiments; and (4) collection of the results and considerations in a validation report [ 5 ]. The validation protocol should clearly define the roles and the responsibilities of each person and element involved in the validation performance, such as equipment, supplies, reagents, reference materials and standards and, above all, the validation parameters and the acceptance criteria that guarantee the fulfillment of the validation specifications.…”

Potency Analysis of Mesenchymal Stromal Cells Using a Combinatorial Assay Matrix Approach

“…However, for advanced-phase clinical trials, regulatory authorities mandate the development of potency assays as part of the release criteria (de Wolf et al, 2017). The challenges of developing potency assays for MSC-like products include the variability of tissue sources, largely undefined mechanisms of action in humans, and lack of reference standards (Bianco et al, 2013; Deskins et al, 2013; Galipeau and Krampera, 2015; Hematti, 2016; Krampera et al, 2013; Ménard and Tarte, 2013; Radrizzani et al, 2016a, 2016b). In addition, analyzing a single effector pathway as a surrogate measure of potency might be misleading, considering that MSCs deploy a plurality of immunemodulatory and regenerative properties that are induced upon interaction with host microenvironmental and immune status cues (Bernardo and Fibbe, 2013; Keating, 2012; Shi et al, 2012).…”

Preclinical Study of Human Bone Marrow-Derived Mesenchymal Stem Cells Using a 3-Dimensional Manufacturing Setting for Enhancing Spinal Fusion

“…Detailed specifications of the physical, biochemical, and therapeutic properties of MSC spheroids were obtained from our preclinical study, with which a framework of the QC standards for MSC spheroids was proposed in this study ( Supplementary Table S3 ), in regard to the identity, purity, viability, and potency within an appropriate limit, range, or distribution, as per the guidelines preliminarily established by global regulatory bodies for licensed MSC drugs. 98 , 99 The QC standards for the identity, purity, and viability could be suggested with reference to the existing guidelines for MSC therapies. However, the therapeutic activity or intended biological effect is often discriminated depending on the individual product attributes so that the acceptability of potency assay methods and standards should be reviewed on a case-by-case basis.…”