Membrane emulsification for the production of uniform poly-N-isopropylacrylamide-coated alginate particles using internal gelation

BACKGROUNDHuman mesenchymal stem/stromal cells (hMSCs) are at the forefront of regenerative medicine applications due to their relatively easy isolation and availability in adults, potential to differentiate and to secrete a range of trophic factors that could determine specialised tissue regeneration. To date, hMSCs have been successfully cultured in vitro on substrates such as polystyrene dishes (TCPS) or microcarriers. However, hMSC sub‐cultivation and harvest typically employs proteolytic enzymes that act by cleaving important cell membrane proteins resulting in long‐term cell damage. In a process where the cells themselves are the product, a non‐enzymatic and non‐damaging harvesting approach is desirable.RESULTSAn alternative system for hMSC expansion and subsequent non‐enzymatic harvest was investigated here. A liquid/liquid two‐phase system was proposed, comprising a selected perfluorocarbon (FC40) and growth medium (DMEM). The cells exhibited similar cell morphologies compared with TCPS. Moreover, they retained their identity and differentiation potential post‐expansion and post‐harvest. Further, no significant difference was found when culturing hMSCs in the culture systems prepared with either fresh or recycled FC40 perfluorocarbon.CONCLUSIONSThese findings make the FC40/DMEM system an attractive alternative for traditional cell culture substrates due to their ease of cell recovery and recyclability, the latter impacting on overall process costs. © 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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“…enzymes, chelators), magnetic or even stimuli‐responsive (e.g. thermo‐responsive, pH‐responsive, electro‐responsive, photo‐responsive).…”

Section: Introductionmentioning

confidence: 99%

Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

Hanga

Murasiewicz

Pacek

et al. 2017

J. Chem. Technol. Biotechnol

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“…Therefore, in this study, NIPAm was copolymerized with hydrophilic AAc or AAm at different ratios to prepare a thermo‐responsive PNIPAm‐ co ‐AAc with optimized LCST value near to body temperature. It was synthesized by RAFT polymerization which is important for getting the desired polymers owing to a higher control on the final molecular characteristics, like polydispersity and molecular weight (Scheme 2a) (García‐Peñas et al, 2019). The successful polymerization of the copolymer was confirmed using 1 H NMR spectroscopy and GPC (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Preparation of thermosensitive PNIPAm‐based copolymer coated cytodex 3 microcarriers for efficient nonenzymatic cell harvesting during 3D culturing

Darge

Chuang

Lai

et al. 2021

Biotech & Bioengineering

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Enzymatic detachment of cells might damage important features and functions of cells and could affect subsequent cell‐based applications. Therefore, nonenzymatic cell detachment using thermosensitive polymer matrix is necessary for maintaining cell quality after harvesting. In this study, we prepared thermosensitive PNIPAm‐co‐AAc‐b‐PS and PNIPAm‐co‐AAm‐b‐PS copolymers and low critical solution temperature (LCST) was tuned near to body temperature. Then, spin coated polymer films were prepared for cell adhesion and thermal‐induced cell detachment. The alpha‐step analysis and scanning electron microscope image of the films suggested that the thickness of the films depends on the molecular weight and concentration which ranged from 206 to 1330 nm for PNIPAm‐co‐AAc‐b‐PS and 97.5–497 nm for PNIPAm‐co‐AAm‐b‐PS. The contact angles of the films verified that the polymer surface was moderately hydrophilic at 37°C. Importantly, RAW264.7 cells were convincingly proliferated on the films to a confluent of >80% within 48 h and abled to detach by reducing the temperature. However, relatively more cells were grown on PNIPAm‐co‐AAm‐b‐PS (5%w/v) films and thermal‐induced cell detachment was more abundant in this formulation. As a result, PNIPAm‐co‐AAm‐b‐PS (5%w/v) was further used to coat commercial cytodex 3 microcarriers for 3D cell culturing and interestingly enhanced cell detachment with preserved potential of recovery was observed at a temperature of below LCST. Thus, surface modification of microcarriers with thermosensitive PNIPAm‐co‐AAm‐b‐PS could be vital strategy for nonenzymatic cell detachment and to achieve adequate number of cells with maximum cell viability and functionality.

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“…The size of CaCO 3 particles should be about 10 times smaller than the pore size of the membrane to prevent pore blockage. A 10-μm microengineered membrane was blocked when the size of CaCO 3 particles suspended in the oil phase phase was 2.3 μm, but no blockage was observed when a 20-μm membrane was used (Hanga and Holdich, 2014). For SPG membrane, blockage is more likely due to tortuous and interconnected pore structure.…”

Section: Internal Gelationmentioning

confidence: 98%

Structured microparticles with tailored properties produced by membrane emulsification

Vladisavljević

2015

Advances in Colloid and Interface Science

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This paper provides an overview of membrane emulsification routes for fabrication of structured microparticles with tailored properties for specific applications. Direct (bottom-up) and premix (top-down) membrane emulsification processes are discussed including operational, formulation and membrane factors that control the droplet size and droplet generation regimes. A special emphasis was put on different methods of controlled shear generation on membrane surface, such as cross flow on the membrane surface, swirl flow, forward and backward flow pulsations in the continuous phase and membrane oscillations and rotations. Droplets produced by membrane emulsification can be used for synthesis of particles with versatile morphology (solid and hollow, matrix and core/shell, spherical and non-spherical, porous and coherent, composite and homogeneous), which can be surface functionalised and coated or loaded with macromolecules, nanoparticles, quantum dots, drugs, phase change materials and high molecular weight gases to achieve controlled/targeted drug release and impart special optical, chemical, electrical, acoustic, thermal and magnetic properties. The template emulsions including metal-in-oil, solid-in-oil-in-water, oil-in-oil, multilayer, and Pickering emulsions can be produced with high encapsulation efficiency of encapsulated materials and narrow size distribution and transformed into structured particles using a variety of different processes, such as polymerisation (suspension, mini-emulsion, interfacial and in-situ), ionic gelation, chemical crosslinking, melt solidification, internal phase separation, layer-by-layer electrostatic deposition, particle self-assembly, complex coacervation, spray drying, sol-gel processing, and molecular imprinting. Particles fabricated from droplets produced by membrane emulsification include nanoclusters, colloidosomes, carbon aerogel particles, nanoshells, polymeric (molecularly imprinted, hypercrosslinked, Janus and core/shell) particles, solder metal powders and inorganic particles. Membrane 2 emulsification devices operate under constant temperature due to low shear rates on the membrane surface, which range from (1−10) × 10 3 s −1 in a direct process to (1−10) × 10 4 s −1 in a premix process.Keywords: Membrane Emulsification; Polymeric microsphere; Microgel; Janus Particle; Core/Shell Particle, Colloidosome. Membrane emulsificationMembrane emulsification (ME) involves preparation of emulsions by pressing a pure dispersed phase or pre-emulsified mixture of the dispersed and continuous phase through a microporous membrane under controlled injection rate and shear conditions. In direct membrane emulsification (DME), one liquid (a dispersed phase) is injected through a microporous membrane into another immiscible liquid (the continuous phase) (Nakashima et al., 1991;, which leads to the formation of droplets at the membrane/continuous phase interface ( Figure 1a). In premix membrane emulsification (PME) (Figure 1b), a pre-emulsion is pressed through the membrane (...

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Membrane emulsification for the production of uniform poly-N-isopropylacrylamide-coated alginate particles using internal gelation

Cited by 20 publications

References 37 publications

Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

Preparation of thermosensitive PNIPAm‐based copolymer coated cytodex 3 microcarriers for efficient nonenzymatic cell harvesting during 3D culturing

Structured microparticles with tailored properties produced by membrane emulsification

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