Biotechnology Letters

, Volume 40, Issue 2, pp 427–435 | Cite as

Cell sheet-based multilayered liver tumor models for anti-cancer drug screening

  • Jianing Yang
  • Shengjun Zhao
  • Yunfei Ji
  • Lili Zhao
  • Qingzhu Kong
  • Qing Zhang
Original Research Paper



To fabricate in vitro cell-dense, three-dimensional (3D) tumor models by employing a cell sheet technology for testing anti-cancer drug efficacy.


The stratified liver tumor models were fabricated by stacking contiguous HepG2 cell sheets. Triple-layer (3L), double-layer (2L), single-layer (1L) cell sheet-based liver tumor models (CSLTMs) demonstrated 106, 96, 85% cell viability, respectively, after 3 days treatment of 10 µM doxorubicin hydrochloride (DOX), while cell viability in two-dimensional (2D) conventional culture (control) was 27%. After 7 days of DOX treatment, the viabilities of 3L, 2L, 1L, control were 24, 14, 3 and 4%, respectively. Probable explanations were blocked diffusion of DOX by the intact and multilayered structure and also hypoxia in the bottom of multilayered cell sheets.


CSLTMs showed a thickness-dependent cytotoxic efficacy of DOX and greater drug resistance than the control, thereby providing useful information toward the development of improved biomimetic tumor models.


Anti-cancer drug Cell sheet technology Liver tumor model Multilayer Three-dimensional culture 



The authors would like to thank the funding support from the Affiliated Hospital of Chengde Medical University.

Supporting Information

Supplementary Fig. 1—HepG2 cells cultured on the surface of the UpCell.

Supplementary Fig. 2—HepG2 cell sheet harvested on (Day -2) had broken up into pieces.

Compliance with ethical standards

Conflict of interest

None of the authors has any relationship with industry or financial associations that might pose a conflict of interest in connection with this work.

Supplementary material

10529_2017_2476_MOESM1_ESM.docx (339 kb)
Supplementary material 1 (DOCX 339 kb)


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  • Jianing Yang
    • 1
  • Shengjun Zhao
    • 2
  • Yunfei Ji
    • 3
  • Lili Zhao
    • 1
  • Qingzhu Kong
    • 1
  • Qing Zhang
    • 4
  1. 1.Department of Spine SurgeryAffiliated Hospital of Chengde Medical UniversityChengdeChina
  2. 2.Department of OrthopedicsChengde Central HospitalChengdeChina
  3. 3.Department of Critical Care MedicineChengde Central HospitalChengdeChina
  4. 4.Department of Respiration MedicineAffiliated Hospital of Chengde Medical UniversityChengdeChina

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