Table 4

Proposed experiments design for evaluation the potential therapeutic mechanism of UC-MSCs in the treatment of COPD

Proposed experimentsCriteriaCell type/methodExpected outcomes
UC-MSC characterisationMSC marker analysisUC-MSCs/flow cytometryMeet ISCT guideline
Differentiation potentialUC-MSCs/In vitro differentiation using commercial kits.Adipogenic, chondrogenic, and osteogenic differentiation
KaryotypeUC-MSCs/G-banding methodNormal post-expansion
Growth factor, cytokines secretionUC-MSCs/ProcartaPlex ImmunoassaysDetection of cytokines and growth factors involves in anti-inflammatory and tissue regeneration process.
Metabolic evaluationMitochondrial activitiesUC-MSCs/agilent seahorse XF cell mito stress testMeasurement of mitochondrial activities of UC-MSCs pre-administration
GlycolysisUC-MSCs/agilent seahorse XF glycolysis stress testMeasurement of glycolysis process of UC-MSC pre-administration
Immunoregulatory assessmentLymphocyte Proliferation AssayUC-MSCs+peripheral mononuclear cells from healthy donorsUC-MSCs inhibit the proliferation rate of lymphocytes in the present of PHA.
UC-MSCs+peripheral mononuclear cells from patients with COPDUC-MSCs inhibit the proliferation rate of lymphocytes in the present of PHA in a similar manner to healthy donor counterpart
Growth factors and cytokines analysisUC-MSC secretion profiles under xeno-free and serum-free culture conditionsUC-MSCs/procartaPlex immunoassaysDetection of cytokines and growth factors involves in anti-inflammatory and tissue regeneration process.
Cytokine profiles from patients with COPD plasmaPatients with COPD plasma/procartaPlex immunoassaysEvaluation of cytokines involves in inflammatory response obtain from patients with COPD plasma before and after UC-MSC administration.
Interaction between UC-MSCs and patients with COPD’ lymphocytesMedia obtained from co-culture of UC-MSCs and peripheral mononuclear cells from patients with COPD/ProcartaPlex ImmunoassaysDetection of cytokines involves in the anti-inflammatory functions of UC-MSCs.

  • COPD, chronic obstructive pulmonary disease; GOLD, Global Initiative for Chronic Obstructive Lung Disease; ISCT, International Society for Cellular Therapy.