Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells

2D surfaces such as tissue culture polystyrene dish used in conventional cell culture have limited surface area for cell growth. During harvest, mechanical scraping or trypsin are used to remove confluent cells from their growing surface, which can be harmful to cells and reduce cell yield. Therefor...

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Main Author: Yong, Hsin Nam Ernest
Format: Thesis (University of Nottingham only)
Language:English
Published: 2023
Subjects:
Online Access:https://eprints.nottingham.ac.uk/72142/
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author Yong, Hsin Nam Ernest
author_facet Yong, Hsin Nam Ernest
author_sort Yong, Hsin Nam Ernest
building Nottingham Research Data Repository
collection Online Access
description 2D surfaces such as tissue culture polystyrene dish used in conventional cell culture have limited surface area for cell growth. During harvest, mechanical scraping or trypsin are used to remove confluent cells from their growing surface, which can be harmful to cells and reduce cell yield. Therefore, there is a need for an improved cell growth surface with increased surface area and an alternative non-invasive cell harvesting mechanism. To this end, a potential three-dimensional (3D) cell culture platform was developed with nanofibers of poly(N-isopropylacrylamide) (PNIPAm) as the main structural framework, fabricated via the electrospinning technique. Collagen type I extracted from fish scales of Tilapia fish was used to functionalize PNIPAm nanofibers scaffold via dip-coating to improve cell-scaffold interaction. The developed electrospun PNIPAm nanofibers scaffold exhibits a morphology that highly mimics the native extracellular matrix (ECM) in terms of fiber diameter, measuring 436.35 ± 187.04 nm. Pore size and porosity were computed as 1.24 ± 1.27 nm and 63.6 %, respectively. Collagen functionalized PNIPAm nanofibers scaffolds show significant improvement in cell viability of rat amniotic fluid stem cells (R3- AFSC) with increasing collagen content, with 50, 70 and 180 % cell viability on neat electrospun PNIPAm, PNIPAm dip-coated in 0.1 and 0.5 % (w/w) collagen solution, respectively. On the other hand, increased collagen content reduces the capability of PNIPAm nanofibers scaffold to spontaneously release R3-AFSC upon cooling from cell culture conditions (37 °C and 5 % CO2) to 25 °C (room temperature), with 51, 48 and 43 % of R3-AFSC released from neat electrospun PNIPAm, PNIPAm dip-coated in 0.1 and 0.5 % (w/w) collagen solution, respectively.
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institution University of Nottingham Malaysia Campus
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spelling nottingham-721422023-12-29T04:30:18Z https://eprints.nottingham.ac.uk/72142/ Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells Yong, Hsin Nam Ernest 2D surfaces such as tissue culture polystyrene dish used in conventional cell culture have limited surface area for cell growth. During harvest, mechanical scraping or trypsin are used to remove confluent cells from their growing surface, which can be harmful to cells and reduce cell yield. Therefore, there is a need for an improved cell growth surface with increased surface area and an alternative non-invasive cell harvesting mechanism. To this end, a potential three-dimensional (3D) cell culture platform was developed with nanofibers of poly(N-isopropylacrylamide) (PNIPAm) as the main structural framework, fabricated via the electrospinning technique. Collagen type I extracted from fish scales of Tilapia fish was used to functionalize PNIPAm nanofibers scaffold via dip-coating to improve cell-scaffold interaction. The developed electrospun PNIPAm nanofibers scaffold exhibits a morphology that highly mimics the native extracellular matrix (ECM) in terms of fiber diameter, measuring 436.35 ± 187.04 nm. Pore size and porosity were computed as 1.24 ± 1.27 nm and 63.6 %, respectively. Collagen functionalized PNIPAm nanofibers scaffolds show significant improvement in cell viability of rat amniotic fluid stem cells (R3- AFSC) with increasing collagen content, with 50, 70 and 180 % cell viability on neat electrospun PNIPAm, PNIPAm dip-coated in 0.1 and 0.5 % (w/w) collagen solution, respectively. On the other hand, increased collagen content reduces the capability of PNIPAm nanofibers scaffold to spontaneously release R3-AFSC upon cooling from cell culture conditions (37 °C and 5 % CO2) to 25 °C (room temperature), with 51, 48 and 43 % of R3-AFSC released from neat electrospun PNIPAm, PNIPAm dip-coated in 0.1 and 0.5 % (w/w) collagen solution, respectively. 2023-07-22 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en cc_by https://eprints.nottingham.ac.uk/72142/1/Amended%20Thesis.pdf Yong, Hsin Nam Ernest (2023) Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells. PhD thesis, University of Nottingham. cell culture; stem cells; nanofibers scaffold; electrospun
spellingShingle cell culture; stem cells; nanofibers scaffold; electrospun
Yong, Hsin Nam Ernest
Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells
title Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells
title_full Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells
title_fullStr Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells
title_full_unstemmed Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells
title_short Development of collagen functionalized thermally responsive poly(N-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3D cell culture platform for rat amniotic fluid stem cells
title_sort development of collagen functionalized thermally responsive poly(n-isopropylacrylamide) electrospun nanofibers scaffold potentially as a 3d cell culture platform for rat amniotic fluid stem cells
topic cell culture; stem cells; nanofibers scaffold; electrospun
url https://eprints.nottingham.ac.uk/72142/