Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)

Secretory factors of adipose-derived stem cells have paracrine effects that may have potential in accelerating proliferation of keratinocytes by coculturing both of the cells. This study was conducted to evaluate the indirect effects of ASCs-PCS on the growth of keratinocytes by paracrine activiti...

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Main Author: Halim, Ahmad Sukari
Format: Monograph
Language:English
Published: Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia 2017
Subjects:
Online Access:http://eprints.usm.my/57504/
http://eprints.usm.my/57504/1/Prof%20Dr%20Ahmad%20Sukari%20Halim-Eprints.pdf
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author Halim, Ahmad Sukari
author_facet Halim, Ahmad Sukari
author_sort Halim, Ahmad Sukari
building USM Institutional Repository
collection Online Access
description Secretory factors of adipose-derived stem cells have paracrine effects that may have potential in accelerating proliferation of keratinocytes by coculturing both of the cells. This study was conducted to evaluate the indirect effects of ASCs-PCS on the growth of keratinocytes by paracrine activities. Human adipose-derived stem cells and human epidermal keratinocytes were isolated and verified with their specific markers. Distribution and proliferation of ASCs on porous chitosan scaffold were assessed by scanning electron microscope, live/dead assay and Alamar blue assay. Coculture groups were divided into two groups; culture ASCs and culture of ASCs-PCS. Proliferation of keratinocytes in the coculture wells were analysed by Alamar blue assay at 24 hours and 72 hours of coculture while the concentration human epidermal growth factor in culture supernatant were quantified at 72 hours of coculture. Adipose-derived stem cells culture have been established and displayed high expression of CD29, CD73, CD90, CD105 and low expression of CD34. Keratinocytes culture expressed involucrin and cytokeratin 6 as demonstrated by immunocytochemistry. Percentage of proliferation of ASCs within PCS was more than 80% accompanied with spindle-shaped features and good viability. After 72 hours of coculture, the percentage of nHEK in the coculture group was increased due to growth medium. ELISA assay revealed no secretion of EGF from ASCs. This study demonstrated via dual -chamber coculture model, human -derived ASCs and PCS showed no growth stimulatory effect towards the growth of keratinocytes via the tested growth factor, EGF. The proliferation of nHEK in the coculture wells possibly due to other growth factor or other mechanism.
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language English
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spelling usm-575042023-03-26T00:11:42Z http://eprints.usm.my/57504/ Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC) Halim, Ahmad Sukari QH573-671 Cytology Secretory factors of adipose-derived stem cells have paracrine effects that may have potential in accelerating proliferation of keratinocytes by coculturing both of the cells. This study was conducted to evaluate the indirect effects of ASCs-PCS on the growth of keratinocytes by paracrine activities. Human adipose-derived stem cells and human epidermal keratinocytes were isolated and verified with their specific markers. Distribution and proliferation of ASCs on porous chitosan scaffold were assessed by scanning electron microscope, live/dead assay and Alamar blue assay. Coculture groups were divided into two groups; culture ASCs and culture of ASCs-PCS. Proliferation of keratinocytes in the coculture wells were analysed by Alamar blue assay at 24 hours and 72 hours of coculture while the concentration human epidermal growth factor in culture supernatant were quantified at 72 hours of coculture. Adipose-derived stem cells culture have been established and displayed high expression of CD29, CD73, CD90, CD105 and low expression of CD34. Keratinocytes culture expressed involucrin and cytokeratin 6 as demonstrated by immunocytochemistry. Percentage of proliferation of ASCs within PCS was more than 80% accompanied with spindle-shaped features and good viability. After 72 hours of coculture, the percentage of nHEK in the coculture group was increased due to growth medium. ELISA assay revealed no secretion of EGF from ASCs. This study demonstrated via dual -chamber coculture model, human -derived ASCs and PCS showed no growth stimulatory effect towards the growth of keratinocytes via the tested growth factor, EGF. The proliferation of nHEK in the coculture wells possibly due to other growth factor or other mechanism. Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia 2017 Monograph NonPeerReviewed application/pdf en http://eprints.usm.my/57504/1/Prof%20Dr%20Ahmad%20Sukari%20Halim-Eprints.pdf Halim, Ahmad Sukari (2017) Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC). Project Report. Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia. (Submitted)
spellingShingle QH573-671 Cytology
Halim, Ahmad Sukari
Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)
title Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)
title_full Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)
title_fullStr Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)
title_full_unstemmed Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)
title_short Involvement of MAPK/ERK and Akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (ACSS)-Porous Chitosan Scaffold (PSC)
title_sort involvement of mapk/erk and akt signaling pathways in the proliferation of keratinocytes cocultured with adipose-derived stem cells (acss)-porous chitosan scaffold (psc)
topic QH573-671 Cytology
url http://eprints.usm.my/57504/
http://eprints.usm.my/57504/1/Prof%20Dr%20Ahmad%20Sukari%20Halim-Eprints.pdf