Hematopoietic Stem and Progenitor Cell Expansion in Contact with Mesenchymal Stromal Cells in a Hanging Drop Model Uncovers Disadvantages of 3D Culture

Figure 5

HSPCs proliferate more extensively in coculture with MSC monolayers than with MSC spheroids. Expansion of HSPCs under 2D or 3D conditions was compared by seeding 500 HSPCs in combination with 5000 MSCs in conventional 96-well culture plates or in hanging drop plates, respectively, followed by determination of HSPC numbers at different time points. (a) Representative data from one donor show a significantly lower proliferation rate already after one week of coculture in 3D when compared with 2D. Samples were analyzed in triplicate. Statistical analysis was performed using a two-tailed parametric -test. (b) The fold increase in HSPC numbers was obtained after expansion of 500 HSPCs in coculture or in monoculture. The highest proliferation rates were detected for the 2D coculture, whereas HSPCs incubated with MSC spheroids in 3D did not exceed the expansion level of the 2D monoculture. Statistical significance was assessed using one-way ANOVA. Data are shown as mean ± standard deviation of four independent experiments with four donors (; ).