Decellularization Techniques to Manufacture Ovary Cancer Niche Scaffold as Prototype for 3D Cancer Cell Culture

DOI

10.2991/978-94-6463-112-8_10How to use a DOI?

Keywords

ovarian cancer; scaffold; decellularization; freeze-thawing; repeated cycles

Abstract

Ovarian cancer contains a microenvironment that supports the proliferation of cancer cells. The microenvironment of ovarian cancer can be produced in vitro by the manufacture of an ovarian cancer scaffold. The scaffold provides conditions structurally similar to the in vivo microenvironment for communication between cells and cells with the extracellular matrix (ECM) thus providing in vitro platform for disease modeling or drug screening. The decellularization method is used to remove cellular components from the ECM with little damage to the structure. The manufacture of ovarian cancer scaffolds in this study are carried out by physical and chemical decellularization techniques with comparison between one, two or three cycles of freeze-thawing. The process of making an ovarian cancer scaffold is using NaOH 0.4N with one, two or three times of freeze-thawing (−80 ℃ and 37 ℃) followed by agitation with an orbital shaker. Characterization of the scaffold was performed by measurement of DNA concentration, histology analysis of decellularization by hematoxylin eosin and scanning electron microscopy (SEM), ECM analysis by masson trichrome for collagen area fraction, collagen III immunohistochemistry, fibronectin immunohistochemistry and byglican immunohistochemistry, cytotoxicity assay of SKOV-3 cell line using flow cytometry 7-AAD. The average genetic material of DNA from the scaffold is lowest at 4.1 ng/μl from the three times freeze-thawed cycles. Hematoxylin-Eosin staining showed a minimum amount of absence of nuclei and presence of intact ECM structure in fibers form particularly in the three times freeze-thawed cycles. SEM result confirmed absence of nuclei and preservation of ECM fibrous structure at the surface of the scaffold in the three times freeze-thawed cycles. The integrity of ECM is preserved with minimal damage to the ECM showed in three times freeze-thawed cycles. Masson’s Trichrome staining showed the collagen fibers in ECM had less damage and more collagen consistently in the three times freeze-thawed cycles. Immunohistochemistry of the ovary cancer scaffold showed the presence of collagen type III, fibronectin, and biglycan with consistently similar area fraction between three times freeze-thawed cycles and non decellularized ovarian cancer tissue which indicate preservation of those ECM proteins. The ovary cancer scaffold displayed very low toxicity to SKOV-3 cell line. The ovarian cancer scaffold manufactured in the three times freeze-thawed cycles has the least DNA, more intact collagen fibers, preserved ECM proteins e.g. collagen type III, fibronectin, and biglycan, and very low toxicity to SKOV-3 cell line.

Copyright

© 2023 The Author(s)

Open Access

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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TY  - CONF
AU  - Siti M. G. Maya
AU  - Nurhamitha Humairoh
AU  - Muhammad Suhaeri
AU  - Farah Nurlidar
AU  - Septelia I. Wanandi
AU  - Tricia D. Anggraeni
AU  - Radiana D. Antarianto
PY  - 2023
DA  - 2023/03/01
TI  - Decellularization Techniques to Manufacture Ovary Cancer Niche Scaffold as Prototype for 3D Cancer Cell Culture
BT  - Proceedings of the 1st International Conference for Health Research – BRIN (ICHR 2022)
PB  - Atlantis Press
SP  - 92
EP  - 104
SN  - 2468-5739
UR  - https://doi.org/10.2991/978-94-6463-112-8_10
DO  - 10.2991/978-94-6463-112-8_10
ID  - Maya2023
ER  -