title: High-Throughput Microfluidic Platform for 3D Cultures of Mesenchymal Stem Cells, Towards Engineering Developmental Processes.
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 μm). 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 TGFβ3 (0.1 ng/ml) permissive to hBM-MSCs 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.
authors.affiliation: 1] Department of Electronics, Information and Bioengineering, Politecnico di Milano, Milano, Italy [2] Departments of Surgery and of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland.
authors.forename: Paola
authors.initials: P
authors.lastname: Occhetta
authors.affiliation: Departments of Surgery and of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland.
authors.forename: Matteo
authors.initials: M
authors.lastname: Centola
authors.affiliation: Departments of Surgery and of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Tonnarelli
authors.affiliation: Department of Electronics, Information and Bioengineering, Politecnico di Milano, Milano, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Redaelli
authors.affiliation: Departments of Surgery and of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland.
authors.forename: Ivan
authors.initials: I
authors.lastname: Martin
authors.affiliation: Department of Electronics, Information and Bioengineering, Politecnico di Milano, Milano, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Rasponi
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024043:Collagen Type II / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology; D060509:Wnt3A Protein / Q000494:pharmacology
chemical_list: D024043:Collagen Type II; D053782:Transforming Growth Factor beta3; D060509:Wnt3A Protein; D016222:Fibroblast Growth Factor 2