An in vitro three-dimensional co-culture system for ameloblastoma modelling

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, Zainal Ariff Abdul Rahman, Tsujigiwa, Hidetsugu, Hamada, Mei, Takabatake, Kiyofumi, Nakano, Keisuke, Nagatsuka, Hitoshi, Chong, Huat Siar
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2019
Online Access:http://journalarticle.ukm.my/13892/
http://journalarticle.ukm.my/13892/
http://journalarticle.ukm.my/13892/1/15%20Soo%20Leng%20Lee.pdf
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spelling ukm-138922020-01-10T08:46:24Z http://journalarticle.ukm.my/13892/ An in vitro three-dimensional co-culture system for ameloblastoma modelling Lee, Soo Leng Zainal Ariff Abdul Rahman, Tsujigiwa, Hidetsugu Hamada, Mei Takabatake, Kiyofumi Nakano, Keisuke Nagatsuka, Hitoshi Chong, Huat Siar 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. Immunohistochemistry, 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 morphodifferentiation. 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 2019-08 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/13892/1/15%20Soo%20Leng%20Lee.pdf Lee, Soo Leng and Zainal Ariff Abdul Rahman, and Tsujigiwa, Hidetsugu and Hamada, Mei and Takabatake, Kiyofumi and Nakano, Keisuke and Nagatsuka, Hitoshi and Chong, Huat Siar (2019) An in vitro three-dimensional co-culture system for ameloblastoma modelling. Sains Malaysiana, 48 (8). pp. 1697-1706. ISSN 0126-6039 http://www.ukm.my/jsm/malay_journals/jilid48bil8_2019/KandunganJilid48Bil8_2019.html
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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. Immunohistochemistry, 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 morphodifferentiation. 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
Zainal Ariff Abdul Rahman,
Tsujigiwa, Hidetsugu
Hamada, Mei
Takabatake, Kiyofumi
Nakano, Keisuke
Nagatsuka, Hitoshi
Chong, Huat Siar
spellingShingle Lee, Soo Leng
Zainal Ariff Abdul Rahman,
Tsujigiwa, Hidetsugu
Hamada, Mei
Takabatake, Kiyofumi
Nakano, Keisuke
Nagatsuka, Hitoshi
Chong, Huat Siar
An in vitro three-dimensional co-culture system for ameloblastoma modelling
author_facet Lee, Soo Leng
Zainal Ariff Abdul Rahman,
Tsujigiwa, Hidetsugu
Hamada, Mei
Takabatake, Kiyofumi
Nakano, Keisuke
Nagatsuka, Hitoshi
Chong, Huat Siar
author_sort Lee, Soo Leng
title An in vitro three-dimensional co-culture system for ameloblastoma modelling
title_short An in vitro three-dimensional co-culture system for ameloblastoma modelling
title_full An in vitro three-dimensional co-culture system for ameloblastoma modelling
title_fullStr An in vitro three-dimensional co-culture system for ameloblastoma modelling
title_full_unstemmed An in vitro three-dimensional co-culture system for ameloblastoma modelling
title_sort in vitro three-dimensional co-culture system for ameloblastoma modelling
publisher Penerbit Universiti Kebangsaan Malaysia
publishDate 2019
url http://journalarticle.ukm.my/13892/
http://journalarticle.ukm.my/13892/
http://journalarticle.ukm.my/13892/1/15%20Soo%20Leng%20Lee.pdf
first_indexed 2023-09-18T20:05:52Z
last_indexed 2023-09-18T20:05:52Z
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