Phenotypic, Functional, and Safety Control at Preimplantation Phase of MSC-Based Therapy

Figure 1

(a) Isolation efficiency. Average number of cells isolated by enzymatic method WJ-MSC(E) is initially higher than cells obtained by mechanical method WJ-MSC(M). Furthermore, under 5% O₂, the number of received cells significantly increases in all variants of isolation. The results are presented as mean values of 3 isolations ± SD, . (b) Flow cytometry analysis. Flow cytometry analysis showed in both types of cultures almost identical typical and relatively high expression of specific mesenchymal markers (CD73, CD90, and CD105). Similarly, not more than 1% of WJ-MSC(E) express negative markers (CD34, CD11b, CD19, CD45, and HLA-DR). (c) Immunocytochemical analysis of WJ-MSC(M) and WJ-MSC(E). Expression of the classical mesenchymal markers revealed presence of the discrete but detectable differences for both types of culture at early passages. (d, e) Quantitative analysis of mesenchymal marker genes expression in early/late passages. Relative expression of WJ-MSC(M) at early (2nd-3rd) and late (8th) passages normalized to the reference gene ACTB was compared to WJ-MSC(E) as calibrator group. The results are presented as mean values of 3 isolations ± SD; ; ; .

(a)

(b)

(c)

(d)

(e)