Development of photodegradable gelatin hydrogels and image analysis technique for automatic optical cell separation system based on the cellular morphology in embedding culture

Abstract

Event Abstract Back to Event Development of photodegradable gelatin hydrogels and image analysis technique for automatic optical cell separation system based on the cellular morphology in embedding culture Masato Tamura1, Shinji Sugiura1, Taku Satoh1, Toshiyuki Kanamori1, Shibuta Mayu2, Kei Kanie2, Ryuji Kato2, Hirofumi Matsui3 and Masumi Yanagisawa4 1 National Institute of Advanced Industrial Science and Technology (AIST), Department of Life Science and Biotechnology, Japan 2 Nagoya University, Graduate School of Pharmaceutical Sciences, Japan 3 University of Tsukuba, Faculty of Medicine, Japan 4 Engineering System Co., Ltd., Japan This paper reports novel cell separation strategy and automated cell separation system based on the cellular morphology under 3D culture environment in the photodegradable hydrogel. Recently, we developed gelatin-based photodegradable hydrogels by NHS-activated-ester reaction [1], and applied this hydrogels to optical cell separation [2]. More recently, we developed “click-crosslinkable and photodegradable gelatin hydrogels” [3]. The present study applied this hydrogels to the most promising application, cell separation based on the cellular morphology. The present study includes proof concept of morphology-based cell separation and development of automatic cell separation system. 3D culture environment with a specific extracellular matrix regulates cellular function and phenotype. In addition, cancer cell morphology is alternated depending on its malignancy in 3D culture environment [4]. Recently we developed the predication model of stem cell differentiation by image analysis [5]. In this paper, we introduced this image analysis technique and the features of click-crosslinkable and photodegradable gelatin hydrogels to the automated morphology-based cell separation system in 3D culture environment in the above mentioned photodegradable gelatin hydrogels. Figure 1 shows the schematic procedure for optical cell separation based on the cellular morphology under 3D culture environment in the photodegradable hydrogel. Suspension of cells including heterogeneous population is mixed with pregel solutions and cells are encapsulated in the gelatin-based photodegradable hydrogels (Figure 1a). After the culture in 3D environment (Figure 1b), microscopic images of the cells are captured. The captured images are analyzed to distinguish the target cells from the other cells by using the image analysis algorithm, which we previously developed for analyzing stem cells (Figure 2) [5]. The hydrogels around the target area is irradiated with UV light (Figure 1c). The cells in the irradiated area are collected by automated pipetting system (Figure 1d). We developed automated system for this optical cell separation procedure, including cultivation, image acquisition, image analysis, light irradiation, and pipetting for cell collection (Figure 3). We are currently developing an automated image analysis algorithm to distinguish cancer cells from normal cells under 3D environment. The automated optical cell separation system with image analysis algorithm will be applied to the establishment of novel cancer-cell lines from clinical samples such as biopsy tissue. KAKENHI (14J07186); NEDO (1009004)