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Published Online: 14 July 2022

Spatial Heterogeneity of Invading Glioblastoma Cells Regulated by Paracrine Factors

Publication: Tissue Engineering Part A
Volume 28, Issue Number 13-14


Glioblastoma (GBM) is the most common and lethal type of malignant primary brain tumor in adults. GBM displays heterogeneous tumor cell population comprising glioma-initiating cells (GICs) with stem cell-like characteristics and differentiated glioma cells. During GBM cell invasion into normal brain tissues, which is the hallmark characteristic of GBM, GICs at the invasion front retain stemness, while cells at the tumor core display cellular differentiation. However, the mechanism of cellular differentiation underlying the formation of spatial cellular heterogeneity in GBM remains unknown. In the present study, we first observed spatially heterogeneous GBM cell populations emerged from an isogenic clonal population of GICs during invasion into a 3D collagen hydrogel in a microfluidic device. Specifically, GICs at the invasion front maintained stemness, while trailing cells displayed astrocytic differentiation. The spatial cellular heterogeneity resulted from the difference in cell density between GICs at the invasion front and trailing cells. Trailing GICs at high cell density exhibited astrocytic differentiation through local accumulation of paracrine factors they secreted, while cells at the invasion front of low cell density retained stemness due to the lack of paracrine factors. In addition, we demonstrated that interstitial flow suppressed astrocytic differentiation of trailing GICs by the clearance of paracrine factors. Our findings suggest that intercellular crosstalk between tumor cells is an essential factor in developing the spatial cellular heterogeneity of GBM cells with various differentiation statuses. It also provides insights into the development of novel therapeutic strategies targeting GBM cells with stem cell characteristics at the invasion front.

Impact Statement

We elucidated the mechanism of cellular differentiation underlying the spatial cellular heterogeneity of glioblastoma composed of glioma-initiating cells (GICs) and differentiated glioma cells during invasion in a microfluidic device. Trailing cells at high cell density exhibited astrocytic differentiation through local accumulation of paracrine factors they produced, while cells at the invasion front of low cell density were shown to retain stemness due to the lack of paracrine factors. Our findings provide valuable knowledge for the development of effective therapeutic strategies targeting GICs at the invasion front.

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Information & Authors


Published In

cover image Tissue Engineering Part A
Tissue Engineering Part A
Volume 28Issue Number 13-14July 2022
Pages: 573 - 585
PubMed: 34841881


Published online: 14 July 2022
Published in print: July 2022
Published ahead of print: 24 January 2022
Published ahead of production: 27 November 2021
Accepted: 22 November 2021
Received: 31 August 2021


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School of Integrated Design Engineering, Keio University, Yokohama, Japan.
Tadahiro Yamashita
School of Integrated Design Engineering, Keio University, Yokohama, Japan.
Department of System Design Engineering, Keio University, Yokohama, Japan.
Oltea Sampetrean
Division of Gene Regulation, Institute for Advanced Medical Research, Keio University School of Medicine, Tokyo, Japan.
Hideyuki Saya
Division of Gene Regulation, Institute for Advanced Medical Research, Keio University School of Medicine, Tokyo, Japan.
School of Integrated Design Engineering, Keio University, Yokohama, Japan.
Department of System Design Engineering, Keio University, Yokohama, Japan.


Address correspondence to: Ryo Sudo, PhD, Department of System Design Engineering, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan [email protected]

Disclosure Statement

No competing financial interests exist.

Funding Information

This work was supported by the Japan Society for the Promotion of Science (19J22105 to Yuta Chonan and 19D04452 to Ryo Sudo), and Japan Agency for Medical Research and Development (JP20gm5810026 to Ryo Sudo).

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