Scientific RepoRts | 7: 192 | DOI:10.1038/s41598-017-00087-x
The combination of three-
dimensional and rotary cell culture
system promotes the proliferation
and maintains the dierentiation
potential of rat BMSCs
, Yan Xu
, Zhifeng Xiao
, Yannan Zhao
, Jing Li
, Sufang Han
, Lei Chen
, Ling Wang
, Bing Chen
& Hong Wang
Bone marrow mesenchymal stem cells (BMSCs) are a good candidate for tissue engineering and clinical
application. One of the challenges in its cell therapy is how to quickly obtain an adequate number of
seed cells and meanwhile maintain suitable dierentiation potential. In this study we combined three-
dimensional (3D) collagen porous scaolds with rotary cell culture system (RCCS) (RCCS-3D) to create a
stereoscopic dynamic environment for the amplication of rat BMSCs in vitro. The results revealed that
this RCCS-3D system could enhance BMSCs’ proliferation and colony formation, as well as maintain the
dierentiation potential compared with conventional static two-dimensional (2D) and 3D cell culture
conditions. In addition, high-throughput microarray analysis showed that gene expressions of RCCS-3D
system displayed signicant dierences in cell proliferation and dierentiation compared with static-2D
conditions. Thus, RCCS-3D system could provide an eective means for BMSCs cell proliferation in vitro
and meanwhile maintain dierentiation potential in tissue engineering.
Bone marrow mesenchymal stem cells (BMSCs) are ideal seed cells for tissue engineering and regenerative med-
icine due to their great properties of self-renewal and pluripotency. Moreover, autogenous BMSCs do not have
exogenous gene safety issues and ethical problems involved in induced pluripotent stem (iPS) cells
onic stem (ES) cells
. However, it’s still a challenge in clinical application of tissue-engineering to obtain su-
cient quantities of BMSCs through static two-dimensional (2-D) expansion in vitro
. What is worse, long-term
sequential cell passaging results in loss of self-renewal and pluripotency in vitro
The cells growing space environment and the dynamic environment can influence the seed cell fate
Compared with conventional 2D cell culture, 3D cell culture has received wide attention for it could mimic a
native cellular 3D niche in vivo to some extent, which is benecial to the exchanges of nutrition and metabolism,
extracellular matrix synthesis and forming of intricate cell-cell and cell-matrix interactions
. Extracellular matrix
(ECM) plays a critical role in cell proliferation and dierentiation in static condition in vitro
. Collagen, the
main component of ECM in vivo, is a major component of ECM and comprises the major (80–90%) organic
Department of Spine Surgery, First Affiliated Hospital of Dalian Medical University, Dalian, 116011, People’s
Republic of China.
Department of Spine Surgery, Sichuan Provincial Orthopedic Hospital, Chengdu, 610041,
People’s Republic of China.
Department of Spine Surgery, Dali Bai Autonomous Prefecture People’s Hospital,
Dali, 671000, People’s Republic of China.
State Key Laboratory of Molecular Developmental Biology, Institute of
Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100190, People’s Republic of China.
Institute of Combined Injury, State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing Engineering
Research Center for Nanomedicine, College of Preventive Medicine, Third Military Medical University, Chongqing,
400038, People’s Republic of China.
Department of Biotechnology, College of Life Sciences, Shandong Agricultural
University, Taian, 271018, People’s Republic of China.
Department of Oncology, First Aliated Hospital of Dalian
Medical University, Dalian, 116011, People’s Republic of China. Yilong Tang and Yan Xu contributed equally to this
work. Correspondence and requests for materials should be addressed to L.W. (email: email@example.com) or B.C.
(email: firstname.lastname@example.org) or H.W. (email: email@example.com)
Received: 1 December 2015
Accepted: 23 January 2017
Published: xx xx xxxx