Int J Thyroidol.  2018 Nov;11(2):160-166. 10.11106/ijt.2018.11.2.160.

Development of the Three-Dimensional Perfusion Culture Technology for the Salivary Ductal Cells

• Kim JW
• Kim JM
• Choi JS

• KMID: 2448987
• DOI: http://doi.org/10.11106/ijt.2018.11.2.160

Abstract

BACKGROUND AND OBJECTIVES
Salivary hypofunction is one of the common side effects after radioiodine therapy, and its pathophysiology is salivary ductal stenosis resulting from ductal cell injury. This study aimed to develop the functional culture environment of human parotid gland ductal cells in in vitro three-dimensional perfusion culture system.
MATERIALS AND METHODS
We compared plastic dish culture method and three-dimensional culture system containing Matrigel and nanofiber. Morphogenesis of reconstituted salivary structures was assessed by histomorphometry. Functional characteristics were assessed by immunohistochemistry and reverse transcription polymerase chain reaction (aquaporin 5, CK7, CK18, connexin 43, and p21). In addition, we designed the media perfusion culture system and identified higher rate of cell proliferation and expression of connexin 43 in perfusion system comparing to dish.
RESULTS
Human parotid ductal cells were well proliferated with the ductal cell characters under environment with Matrigel. In the presence of Matrigel, aquaporin 5, CK18 and connexin 43 were more expressed than 2D dish and 3D nanofiber setting. In the media perfusion culture system, ductal cells in 3D culture media showed higher cells count and connexin 43 expression compared to 2D dish.
CONCLUSION
This in vitro ductal cell perfusion culture system using Matrigel could be used to study for radioiodine induced sialadenitis model in vivo.

Keyword

Thyroid cancer; Perfusion; Culture; Salivary gland

MeSH Terms

Aquaporin 5
Cell Proliferation
Connexin 43
Constriction, Pathologic
Culture Media
Humans
Immunohistochemistry
In Vitro Techniques
Methods
Morphogenesis
Nanofibers
Parotid Gland
Perfusion*
Plastics
Polymerase Chain Reaction
Reverse Transcription
Salivary Ducts*
Salivary Glands
Sialadenitis
Thyroid Neoplasms
Aquaporin 5
Connexin 43
Culture Media
Plastics

Figure

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  Fig. 1. A schematic diagram of a three-dimensional culture model of ductal cells (A), photographs of ductal cell perfusion devices (B), and optical microscopic photograph of ductal cell culture (C).

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  Fig. 2. (A) The morphology of salivary ductal cells according to 2-D and 3-D culture conditions, (B) The mRNA expression of acinar cell marker (AQP5), ductal cell markers (CK7,18) and cell junctional protein (connexin 43) (∗p＜0.05, ∗∗ p＜0.01, ∗∗∗p＜0.001).

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  Fig. 3. Conformational change of salivary ductal cells under perfusion culture system of ductal cells (A), confirmation of cell gap junctional protein (connexin 43) expression in two-dimensional culture and perfusion culture (B).

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  Fig. 4. (A, B) Viability of salivary duct cells over time after radioiodine treatment. (C) Expression of p21 over time after radioiodine treatment (∗p＜0.05).

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