Abstract
The aim of the present study was to determine the effects of KGF on the differentiation of cultured human amnion epithelial cells (HAECs) towards skin keratinocyte. HAECs at passage 1 were cultured in medium HAM's F12: Dulbecco's Modified Eagles Medium (1:1) supplemented with different concentrations of KGF (0, 5, 10, 20, 30 and 50 ng/ml KGF). Dose-response of KGF on HAECs was determined by morphological assessment; growth kinetic evaluation; immunocytochemical analysis; stemness and epithelial gene expression quantification with two step real time RT-PCR. KGF promotes the proliferation of HAECs with maximal effect observed at 10 ng/ml KGF. However, KGF decreased the stemness genes expression: Oct-3/4, Sox-2, Nanog3, Rex-1, FGF-4, FZD-9 and BST-1. KGF also down-regulates epithelial genes expression: CK3, CK18, CK19, Integrin-β1, p63 and involucrin in cultured HAECs. No significant difference on the gene expression was detected for each Nestin, ABCG-2, CK1 and CK14 in KGF-treated HAECs. Immunocytochemical analysis for both control and KGF-treated HAECs demonstrated positive staining against CK14 and CK18 but negative staining against involucrin. The results suggested that KGF stimulates an early differentiation of HAECs towards epidermal cells. Differentiation of KGF-treated HAECs to corneal lineage is unfavourable. Therefore, further studies are needed to elucidate the roles of KGF in the differentiation of HAECs towards skin keratinocytes.
| Original language | English |
|---|---|
| Pages (from-to) | 905-915 |
| Number of pages | 11 |
| Journal | Burns |
| Volume | 39 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - Aug 2013 |
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Keywords
- Amnion cells
- Differentiation
- Epithelial
- Keratin
- Keratinocyte growth factor
- Skin
- Stemness
ASJC Scopus subject areas
- Emergency Medicine
- Critical Care and Intensive Care Medicine
- Surgery
Cite this
Effects of keratinocyte growth factor on skin epithelial differentiation of human amnion epithelial cells. / Fatimah, Simat Siti; Tan, Geok Chin; Kien Hui, Chua; Tan, Ay Eeng; Abdul Ghani, Nur Azurah; Hayati, Abdul Rahman.
In: Burns, Vol. 39, No. 5, 08.2013, p. 905-915.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Effects of keratinocyte growth factor on skin epithelial differentiation of human amnion epithelial cells
AU - Fatimah, Simat Siti
AU - Tan, Geok Chin
AU - Kien Hui, Chua
AU - Tan, Ay Eeng
AU - Abdul Ghani, Nur Azurah
AU - Hayati, Abdul Rahman
PY - 2013/8
Y1 - 2013/8
N2 - The aim of the present study was to determine the effects of KGF on the differentiation of cultured human amnion epithelial cells (HAECs) towards skin keratinocyte. HAECs at passage 1 were cultured in medium HAM's F12: Dulbecco's Modified Eagles Medium (1:1) supplemented with different concentrations of KGF (0, 5, 10, 20, 30 and 50 ng/ml KGF). Dose-response of KGF on HAECs was determined by morphological assessment; growth kinetic evaluation; immunocytochemical analysis; stemness and epithelial gene expression quantification with two step real time RT-PCR. KGF promotes the proliferation of HAECs with maximal effect observed at 10 ng/ml KGF. However, KGF decreased the stemness genes expression: Oct-3/4, Sox-2, Nanog3, Rex-1, FGF-4, FZD-9 and BST-1. KGF also down-regulates epithelial genes expression: CK3, CK18, CK19, Integrin-β1, p63 and involucrin in cultured HAECs. No significant difference on the gene expression was detected for each Nestin, ABCG-2, CK1 and CK14 in KGF-treated HAECs. Immunocytochemical analysis for both control and KGF-treated HAECs demonstrated positive staining against CK14 and CK18 but negative staining against involucrin. The results suggested that KGF stimulates an early differentiation of HAECs towards epidermal cells. Differentiation of KGF-treated HAECs to corneal lineage is unfavourable. Therefore, further studies are needed to elucidate the roles of KGF in the differentiation of HAECs towards skin keratinocytes.
AB - The aim of the present study was to determine the effects of KGF on the differentiation of cultured human amnion epithelial cells (HAECs) towards skin keratinocyte. HAECs at passage 1 were cultured in medium HAM's F12: Dulbecco's Modified Eagles Medium (1:1) supplemented with different concentrations of KGF (0, 5, 10, 20, 30 and 50 ng/ml KGF). Dose-response of KGF on HAECs was determined by morphological assessment; growth kinetic evaluation; immunocytochemical analysis; stemness and epithelial gene expression quantification with two step real time RT-PCR. KGF promotes the proliferation of HAECs with maximal effect observed at 10 ng/ml KGF. However, KGF decreased the stemness genes expression: Oct-3/4, Sox-2, Nanog3, Rex-1, FGF-4, FZD-9 and BST-1. KGF also down-regulates epithelial genes expression: CK3, CK18, CK19, Integrin-β1, p63 and involucrin in cultured HAECs. No significant difference on the gene expression was detected for each Nestin, ABCG-2, CK1 and CK14 in KGF-treated HAECs. Immunocytochemical analysis for both control and KGF-treated HAECs demonstrated positive staining against CK14 and CK18 but negative staining against involucrin. The results suggested that KGF stimulates an early differentiation of HAECs towards epidermal cells. Differentiation of KGF-treated HAECs to corneal lineage is unfavourable. Therefore, further studies are needed to elucidate the roles of KGF in the differentiation of HAECs towards skin keratinocytes.
KW - Amnion cells
KW - Differentiation
KW - Epithelial
KW - Keratin
KW - Keratinocyte growth factor
KW - Skin
KW - Stemness
UR - http://www.scopus.com/inward/record.url?scp=84879014238&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84879014238&partnerID=8YFLogxK
U2 - 10.1016/j.burns.2012.10.019
DO - 10.1016/j.burns.2012.10.019
M3 - Article
C2 - 23273814
AN - SCOPUS:84879014238
VL - 39
SP - 905
EP - 915
JO - Burns
JF - Burns
SN - 0305-4179
IS - 5
ER -