Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production

Abstract

Background Cell based therapies, such as bone marrow derived mesenchymal stem cells (BM-MSCs; also known as mesenchymal stromal cells), are currently under investigation for a number of disease applications. The current challenge facing the field is maintaining the consistency and quality of cells especially for cell dose production for pre-clinical testing and clinical trials. Here we determine how BM-donor variability and thus the derived MSCs factor into selection of the optimal primary cell lineage for cell production and testing in a pre-clinical swine model of trauma induced acute respiratory distress syndrome. Methods We harvested bone marrow and generated three different primary BM-MSCs from Yorkshire swine. Cells from these three donors were characterized based on (a) phenotype (morphology, differentiation capacity and flow cytometry), (b) in vitro growth kinetics and metabolic activity, and (c) functional analysis based on inhibition of lung endothelial cell permeability. Results Cells from each swine donor exhibited varied morphology, growth rate, and doubling times. All expressed the same magnitude of standard MSC cell surface markers by flow cytometry and had similar differentiation potential. Metabolic activity and growth potential at each of the passages varied between the three primary cell cultures. More importantly, the functional potency of the MSCs on inhibition of endothelial permeability was also cell donor dependent. Conclusion This study suggests that for production of MSCs for cell-based therapy, it is imperative to examine donor variability and characterize derived MSCs for marker expression, growth and differentiation characteristics and testing potency in application dependent assays prior to selection of the optimal cell lineage for large scale expansion and dose production. Electronic supplementary material The online version of this article (10.1186/s12967-019-1877-4) contains supplementary material, which is available to authorized users.

Endometrial stromal cells exhibit a distinct phenotypic and immunomodulatory profile

“…As previously reported, eSCs showed multipotent differentiation potential; however, small variations/ predispositions in the degree of differentiation were observed between donors. In line with existing knowledge of the stroma being composed of heterogeneous subpopulations, different proliferation rates, cell morphologies, immunophenotype, and multi-differentiation potential can all be expected [38–41]. This is an important consideration in developing cell therapy products and should be taken into consideration with regard to the expected therapeutic mode of action.…”

Multi-pronged approach to human mesenchymal stromal cells senescence quantification with a focus on label-free methods

“…Furthermore, from literature, confluence and different media components are the factors to be considered to affect the autofluorescence signals 36 . In our study, we made sure that the media composition was consistent across all samples and the starting seeding concentration for autofluorescence samples remained the same; however, variability in the samples being measured due to cellular heterogeneity may still exist and is donor related 44 . In addition, the heterogeneity and auto-differentiation characteristic of MSC could have resulted in the differences in fold-changes (between early-passage and senescent MSCs) observed for different cultures.…”

Shattering barriers toward clinically meaningful MSC therapies

“…Standardizing therapeutic potency of the MSC product is crucial before beginning clinical trials (30). This need has prompted research efforts toward developing improved in vitro potency assays that can accurately correlate the CQAs of the MSC product with their therapeutic function (31)(32)(33)(34)(35).…”