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An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)

Ameloblastoma, the most clinically significant odontogenic epithelial tumor, is a locally-invasive and destructive lesion in the jawbones. However, the nature of this infiltrativeness and destructive behavior remains ill-understood. To address this, we established an in vitro three-dimensional (3D)...

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Main Authors: Lee, Soo Leng, Rahman, Zainal Ariff Abdul, Tsujigiwa, Hidetsugu, Hamada, Mei, Takabatake, Kiyofumi, Nakano, Keisuke, Nagatsuka, Hitoshi, Siar, Chong Huat
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2019
Subjects:
R Medicine
RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Online Access:http://eprints.um.edu.my/23888/1/15%20Soo%20Leng%20Lee.pdf
http://eprints.um.edu.my/23888/
http://www.ukm.my/jsm/pdf_files/SM-PDF-48-8-2019/15%20Soo%20Leng%20Lee.pdf
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spelling my.um.eprints.238882020-02-21T04:01:30Z http://eprints.um.edu.my/23888/ An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma) Lee, Soo Leng Rahman, Zainal Ariff Abdul Tsujigiwa, Hidetsugu Hamada, Mei Takabatake, Kiyofumi Nakano, Keisuke Nagatsuka, Hitoshi Siar, Chong Huat R Medicine RC0254 Neoplasms. Tumors. Oncology (including Cancer) Ameloblastoma, the most clinically significant odontogenic epithelial tumor, is a locally-invasive and destructive lesion in the jawbones. However, the nature of this infiltrativeness and destructive behavior remains ill-understood. To address this, we established an in vitro three-dimensional (3D) co-culture system to simulate an amelobastoma disease model aimed at investigating the interactions between tumor cells and osteoblasts. Osteoblastic cell lines (KUSA/A1 and mc3T3-E1) and one stromal cell line (ST2) were separately co-seeded with ameloblastoma-derived cell line (AM-1) in a collagen scaffold (representing the extracellular bone matrix) and incubated with mineralization medium. Inununohistochemistry, double immunofluorescence and mineralization assay were performed. Only Am-1/KUsA-A1 co-culture showed a significant increase in Am-1 cell count, suggesting that heterotypic cell-cell interaction promotes tumoral cell growth, while formation of visible Am-1 epithelial nest-like structures resembling ameloblastoma cells in their native state, suggest morphodffferentiation. A RANK-high, RANKL-low and osteoprotegerin-low immunoprofile in co-culture AM-1 cells implies deregulated osteoclastogenesis. Mineralization assays showed diminished calcification in Am-1/KUSA-A1 co-culture extracellular matrix suggesting an altered local bone metabolism. In contrast, KUSA/A1 monocultures showed abundant extracellular matrix calcification. Taken together, these results suggest that a 3D co-culture system as an amelobastoma disease model provides insights that bidirectional ameloblastoma-osteoblastic interactions might play a role in modulating tumor growth and osteoclastogenesis. Penerbit Universiti Kebangsaan Malaysia 2019 Article PeerReviewed text en http://eprints.um.edu.my/23888/1/15%20Soo%20Leng%20Lee.pdf Lee, Soo Leng and Rahman, Zainal Ariff Abdul and Tsujigiwa, Hidetsugu and Hamada, Mei and Takabatake, Kiyofumi and Nakano, Keisuke and Nagatsuka, Hitoshi and Siar, Chong Huat (2019) An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma). Sains Malaysiana, 48 (8). pp. 1697-1706. ISSN 0126-6039 http://www.ukm.my/jsm/pdf_files/SM-PDF-48-8-2019/15%20Soo%20Leng%20Lee.pdf doi:10.17576/jsm-2019-4808-15
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
language English
topic R Medicine
RC0254 Neoplasms. Tumors. Oncology (including Cancer)
spellingShingle R Medicine
RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Lee, Soo Leng
Rahman, Zainal Ariff Abdul
Tsujigiwa, Hidetsugu
Hamada, Mei
Takabatake, Kiyofumi
Nakano, Keisuke
Nagatsuka, Hitoshi
Siar, Chong Huat
An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)
description Ameloblastoma, the most clinically significant odontogenic epithelial tumor, is a locally-invasive and destructive lesion in the jawbones. However, the nature of this infiltrativeness and destructive behavior remains ill-understood. To address this, we established an in vitro three-dimensional (3D) co-culture system to simulate an amelobastoma disease model aimed at investigating the interactions between tumor cells and osteoblasts. Osteoblastic cell lines (KUSA/A1 and mc3T3-E1) and one stromal cell line (ST2) were separately co-seeded with ameloblastoma-derived cell line (AM-1) in a collagen scaffold (representing the extracellular bone matrix) and incubated with mineralization medium. Inununohistochemistry, double immunofluorescence and mineralization assay were performed. Only Am-1/KUsA-A1 co-culture showed a significant increase in Am-1 cell count, suggesting that heterotypic cell-cell interaction promotes tumoral cell growth, while formation of visible Am-1 epithelial nest-like structures resembling ameloblastoma cells in their native state, suggest morphodffferentiation. A RANK-high, RANKL-low and osteoprotegerin-low immunoprofile in co-culture AM-1 cells implies deregulated osteoclastogenesis. Mineralization assays showed diminished calcification in Am-1/KUSA-A1 co-culture extracellular matrix suggesting an altered local bone metabolism. In contrast, KUSA/A1 monocultures showed abundant extracellular matrix calcification. Taken together, these results suggest that a 3D co-culture system as an amelobastoma disease model provides insights that bidirectional ameloblastoma-osteoblastic interactions might play a role in modulating tumor growth and osteoclastogenesis.
format Article
author Lee, Soo Leng
Rahman, Zainal Ariff Abdul
Tsujigiwa, Hidetsugu
Hamada, Mei
Takabatake, Kiyofumi
Nakano, Keisuke
Nagatsuka, Hitoshi
Siar, Chong Huat
author_facet Lee, Soo Leng
Rahman, Zainal Ariff Abdul
Tsujigiwa, Hidetsugu
Hamada, Mei
Takabatake, Kiyofumi
Nakano, Keisuke
Nagatsuka, Hitoshi
Siar, Chong Huat
author_sort Lee, Soo Leng
title An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)
title_short An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)
title_full An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)
title_fullStr An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)
title_full_unstemmed An in vitro Three-Dimensional Co-Culture System for Ameloblastoma Modelling (Sistem Ko-Kultur Tiga Dimensi secara in vitro untuk Pemodelan Ameloblastoma)
title_sort in vitro three-dimensional co-culture system for ameloblastoma modelling (sistem ko-kultur tiga dimensi secara in vitro untuk pemodelan ameloblastoma)
publisher Penerbit Universiti Kebangsaan Malaysia
publishDate 2019
url http://eprints.um.edu.my/23888/1/15%20Soo%20Leng%20Lee.pdf
http://eprints.um.edu.my/23888/
http://www.ukm.my/jsm/pdf_files/SM-PDF-48-8-2019/15%20Soo%20Leng%20Lee.pdf
_version_ 1662755193261916160
score 13.211869

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