Occhetta, Paola, Centola, Matteo, Tonnarelli, Beatrice, Redaelli, Alberto, Martin, Ivan and Rasponi, Marco (2015) High-Throughput Microfluidic Platform for 3D Cultures of Mesenchymal Stem Cells, Towards Engineering Developmental Processes. Scientific Reports, 5. ISSN 2045-2322

Abstract

The development of in vitro models to screen the effect of different concentrations, combinations
and temporal sequences of morpho-regulatory factors on stem/progenitor cells is crucial to
investigate and possibly recapitulate developmental processes with adult cells. Here, we designed
and validated a microfluidic platform to (i) allow cellular condensation, (ii) culture 3D micromasses
of human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) under continuous
flow perfusion, and (ii) deliver defined concentrations of morphogens to specific culture units.
Condensation of hBM-MSCs was obtained within 3 hours, generating micromasses in uniform sizes
(56.2 ± 3.9 μ m). As compared to traditional macromass pellet cultures, exposure to morphogens
involved in the first phases of embryonic limb development (i.e. Wnt and FGF pathways) yielded
more uniform cell response throughout the 3D structures of perfused micromasses (PMMs), and
a 34-fold higher percentage of proliferating cells at day 7. The use of a logarithmic serial dilution
generator allowed to identify an unexpected concentration of TGFβ 3 (0.1 ng/ml) permissive to hBMMSCs
proliferation and inductive to chondrogenesis. This proof-of-principle study supports the
described microfluidic system as a tool to investigate processes involved in mesenchymal progenitor
cells differentiation, towards a ‘developmental engineering’ approach for skeletal tissue regeneration.

Item Type:	Article
Date Deposited:	26 Apr 2016 23:45
Last Modified:	26 Apr 2016 23:45
URI:	https://oak.novartis.com/id/eprint/26006