Development of a contacting transwell co-culture system for the in vitro propagation of primary central nervous system lymphoma

Mayuko Nishi; Kensuke Tateishi; Jeremiah Stanleyraj Sundararaj; Yoko Ino; Yusuke Nakai; Yasuyoshi Hatayama; Yutaro Yamaoka; Yusaku Mihana; Kei Miyakawa; Hirokazu Kimura; Yayoi Kimura; Tetsuya Yamamoto; Akihide Ryo

doi:10.3389/fcell.2023.1275519

Development of a contacting transwell co-culture system for the in vitro propagation of primary central nervous system lymphoma

Front Cell Dev Biol. 2023 Nov 27:11:1275519. doi: 10.3389/fcell.2023.1275519. eCollection 2023.

Authors

Mayuko Nishi^{1

2}, Kensuke Tateishi^{3

4}, Jeremiah Stanleyraj Sundararaj², Yoko Ino⁵, Yusuke Nakai^{1

2}, Yasuyoshi Hatayama^{1

2}, Yutaro Yamaoka^{2

6}, Yusaku Mihana^{2

6}, Kei Miyakawa^{2

7}, Hirokazu Kimura⁸, Yayoi Kimura⁵, Tetsuya Yamamoto³, Akihide Ryo^{1

2}

Affiliations

¹ Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan.
² Department of Microbiology, Graduate School of Medicine, Yokohama City University, Yokohama, Japan.
³ Department of Neurosurgery, Graduate School of Medicine, Yokohama City University, Yokohama, Japan.
⁴ Laboratory of Biopharmaceutical and Regenerative Science, Graduate School of Medical Science, Yokohama City University, Yokohama, Japan.
⁵ Advanced Medical Research Center, Yokohama City University, Yokohama, Japan.
⁶ Life Science Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Tokyo, Japan.
⁷ Research Center for Influenza and Respiratory Viruses, National Institute of Infectious Diseases, Tokyo, Japan.
⁸ Department of Health Science, Gunma Paz University Graduate School of Health Sciences, Takasaki-shi, Japan.

Abstract

Primary central nervous system lymphoma (PCNSL) is a malignant neoplasm of the central nervous system that is refractory to treatment and has extremely poor prognosis. One factor hindering the development of therapeutic options for PCNSL is its molecular heterogeneity and the extreme difficulty in establishing in vitro cell lines that permit intensive research on this disease. In the present study, we developed a method to propagate PCNSL cells in vitro using a contacting transwell cell culture system involving brain vascular pericytes. The co-culture system was found to recapitulate the tumor microenvironment that is influenced by the biological activity of adjacent pericytes, and to sustain the survival and proliferation of PCNSL cells in vitro. We further delineated the underlying molecular mechanisms and found that the HGF-c-Met axis may be involved in the long-term in vitro culture of PCNSL cells. Moreover, the peptidylprolyl isomerase Pin1 was found to play a key role in PCNSL cell survival and it sustained proliferation through interactions with key transcription factors related to B-cell lymphomagenesis. These results suggest that our in vitro co-culture system is well suited to analyzing the biological and molecular characteristics of PCNSL, and may contribute to the discovery of new therapeutic interventions.

Keywords: HGF; NFAT; NFkB; PCNSL; Pin1; lymphoma; pericytes; vitrigel.

Grants and funding

The authors declare that this study received funding from Kanto Chemical Co., Inc. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication.