Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng
Ameloblastoma, the most clinically significant odontogenic epithelial tumour, is a locally-invasive and destructive lesion in the jawbones. Stromal cells as key contributors to the tumour microenvironment have a prominent role in tumour growth, progression and the spread of tumours. Therefore, th...
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my.um.stud.104112020-02-25T18:43:53Z Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng Lee, Soo Leng RK Dentistry Ameloblastoma, the most clinically significant odontogenic epithelial tumour, is a locally-invasive and destructive lesion in the jawbones. Stromal cells as key contributors to the tumour microenvironment have a prominent role in tumour growth, progression and the spread of tumours. Therefore, the reciprocal parenchymal-stromal interactions in the milieu of the tumour microenvironment are inevitably capable of addressing the illunderstood nature of the infiltrativeness and destructive behaviour of ameloblastoma. Objective: An in vitro three-dimensional (3D) ameloblastoma tumour-osteoblast coculture model was established to elucidate the effect of heterotypic cell interactions on tumour growth and morphological characteristics of tumour cell. Materials and Methods: Stromal cell line, ST2 cells, pre-osteoblastic cell line, KUSA/A1 cells and osteoblastic cell line, MC3T3-E1 cells were separately co-seeded with the ameloblastoma tumour cell line, AM-1 in collagen gel incubated with mineralization medium. Results: AM-1/KUSA-A1 co-culture showed a heterogeneous cell population with two distinct morphologies: elongated spindle-shaped vimentin-positive cells with long anastomosing cytoplasmic processes interspersed and encircled cytokeratin-positive round cell which organized into nest-like aggregates. Both round cell with nest-like structures and elongated spindle-shaped cells in co-culture strongly expressed RANK, mildly for RANKL and OPG. 14-day-old KUSA/A1 monocultures shown evidence of intense extracellular matrix mineralization as confirmed by intense Alizarin Red S staining. In contrast, KUSA/A1 cells in the AM co-culture shown reduced Alizarin Red S staining revealed diminished calcification. Furthermore, 3D AM co-cultures showed a significant increase in AM-1 cell count compared to their monoculture counterparts, and formation of visible AM-1 epithelial nest-like structures resembling ameloblastoma cells in their native state. Conclusion: The in vitro 3D co-culture system as established in the present study provided some insights into the biological behaviour of enigmatic amelobastoma iv disease. Present in vitro findings suggest that, bidirectional ameloblastoma-osteoblastic interactions might play an important role in modulating tumour growth and local bone metabolism. 2018 Thesis NonPeerReviewed application/pdf http://studentsrepo.um.edu.my/10411/4/soo_leng.pdf Lee, Soo Leng (2018) Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng. Masters thesis, University of Malaya. http://studentsrepo.um.edu.my/10411/ |
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RK Dentistry Lee, Soo Leng Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng |
description |
Ameloblastoma, the most clinically significant odontogenic epithelial tumour, is a
locally-invasive and destructive lesion in the jawbones. Stromal cells as key contributors
to the tumour microenvironment have a prominent role in tumour growth, progression
and the spread of tumours. Therefore, the reciprocal parenchymal-stromal interactions in
the milieu of the tumour microenvironment are inevitably capable of addressing the illunderstood nature of the infiltrativeness and destructive behaviour of ameloblastoma.
Objective: An in vitro three-dimensional (3D) ameloblastoma tumour-osteoblast coculture model was established to elucidate the effect of heterotypic cell interactions on
tumour growth and morphological characteristics of tumour cell. Materials and
Methods: Stromal cell line, ST2 cells, pre-osteoblastic cell line, KUSA/A1 cells and
osteoblastic cell line, MC3T3-E1 cells were separately co-seeded with the ameloblastoma
tumour cell line, AM-1 in collagen gel incubated with mineralization medium. Results:
AM-1/KUSA-A1 co-culture showed a heterogeneous cell population with two distinct
morphologies: elongated spindle-shaped vimentin-positive cells with long anastomosing
cytoplasmic processes interspersed and encircled cytokeratin-positive round cell which
organized into nest-like aggregates. Both round cell with nest-like structures and
elongated spindle-shaped cells in co-culture strongly expressed RANK, mildly for
RANKL and OPG. 14-day-old KUSA/A1 monocultures shown evidence of intense
extracellular matrix mineralization as confirmed by intense Alizarin Red S staining. In
contrast, KUSA/A1 cells in the AM co-culture shown reduced Alizarin Red S staining
revealed diminished calcification. Furthermore, 3D AM co-cultures showed a significant
increase in AM-1 cell count compared to their monoculture counterparts, and formation
of visible AM-1 epithelial nest-like structures resembling ameloblastoma cells in their
native state. Conclusion: The in vitro 3D co-culture system as established in the present
study provided some insights into the biological behaviour of enigmatic amelobastoma
iv
disease. Present in vitro findings suggest that, bidirectional ameloblastoma-osteoblastic
interactions might play an important role in modulating tumour growth and local bone
metabolism. |
format |
Thesis |
author |
Lee, Soo Leng |
author_facet |
Lee, Soo Leng |
author_sort |
Lee, Soo Leng |
title |
Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng |
title_short |
Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng |
title_full |
Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng |
title_fullStr |
Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng |
title_full_unstemmed |
Development of an in vitro 3D co-culture system as an ameloblastoma disease model / Lee Soo Leng |
title_sort |
development of an in vitro 3d co-culture system as an ameloblastoma disease model / lee soo leng |
publishDate |
2018 |
url |
http://studentsrepo.um.edu.my/10411/4/soo_leng.pdf http://studentsrepo.um.edu.my/10411/ |
_version_ |
1738506363338752000 |
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13.160551 |