Shape-dependent regulation of differentiation lineages of bone marrow-derived cells under cyclic stretch
Introduction
Stem cells have a multipotent differentiation capability and have been considered as a key material in the success of regenerative medicine. Bone marrow is a primary source of the stem cells among other tissues such as blood and adipose tissue. Bone marrow-derived stromal cells, also named as bone marrow mesenchymal stem cells (bMSCs), have been studied extensively for their multipotent differentiation capability. Moreover, an application of mechanical loading has been demonstrated to be effective in directing differentiation towards cells in load-bearing tissues such as blood vessel (vascular smooth muscle cells), bone (osteoblasts) and articular cartilage (articular chondrocytes) (Steward and Kelly, 2015).
Among a variety of mechanical loading regimens, cyclic stretch has been adopted in a number of studies in an attempt to lead bMSCs into specific lineages. It has been demonstrated that the application of cyclic stretch to bMSCs in vitro resulted in changes in the expression of markers for osteogenic, chondrogenic, smooth muscle and tenogenic differentiations. The direction of the change (i.e., upregulation or downregulation) depends on the amplitude of cyclic stretch. In general, cyclic stretch with a low amplitude up to 5% induces an upregulation of osteogenic differentiation (Byrne et al., 2008, Haudenschild et al., 2009, Kearney et al., 2010, Koike et al., 2005, Qi et al., 2008, Rui et al., 2011, Ward et al., 2007), whereas that with an amplitude over 10% resulted in the enhancement of the differentiation towards smooth muscle cells (Ghazanfari et al., 2009, Jang et al., 2011). Tenogenic differentiation is inducible by cyclic stretch with an amplitude ranging from 1% to 10% (Kuo and Tuan, 2008, Morita et al., 2019, Morita et al., 2013).
It is known that bone marrow contains cells with a variety of shapes. Two types of cell shape were reported first (Mets and Verdonk, 1981): fibroblast-like cells and large, epithelial-like cells. This was followed by a report showing the presence of round-shape cells (Colter et al., 2001, Kobayashi et al., 2004, Vogel et al., 2003). These cell shapes seemed to represent different differentiation capabilities; fibroblast-like spindle cells can be differentiated into smooth muscle cells under fluid flow stimulation, while round-shape cells and large cells can be differentiated into adipocytes and osteoblast, respectively (Kobayashi et al., 2004). Chondrogenic and neurogenic differentiation capabilities have also been confirmed (Freeman et al., 2015, Ward et al., 2007). Despite the presence of such heterogeneity in the population of bone marrow-derived cells, in the most of past studies, cells were isolated from the marrow of long bones, expanded and cultured in tissue culture flasks before the use in subsequent culture experiments for cell differentiation as stem cells (e.g., Both et al., 2007, Farrell et al., 2006). During these procedures, non-adherent cells, which could include hematopoietic cells, were excluded so that the remaining, cells attaching to a plastic substrate were selectively cultured to confluency, at least once, to make them a homogeneous population of stem cells. Although this could potentially give researchers stable and reproducible materials, such stem cells still demonstrate widely varying differentiation potentials between individual cells (Freeman et al., 2015). In addition, the use of stem cell lines comes with a drawback that the differentiation capability of the original, native cells in bone marrow cannot be analyzed. The heterogeneity of bone marrow-derived stem cells is thought to lead to an unsatisfactory outcome of tissue repair by the administration of these cell types in clinical settings (Huang et al., 2010, Wang et al., 2015). For a better outcome in the therapeutic use of stem cells, understanding of how the native bone marrow-derived cells can be differentiated to various specific lineages would be useful, particularly in the presence of extrinsic signals such as mechanical and/or chemical stimuli. Therefore, the present study has been performed to investigate how the differentiation of post-harvest, native bone marrow-derived cells is regulated by cyclic stretch in vitro.
Section snippets
Bone marrow-derived cell isolation
All animal experiments were approved by the institutional review board for animal care at Nagoya Institute of Technology (Approval No. 17003) and were performed following the Guide for Animal Experimentation, Nagoya Institute of Technology. Bone marrow-derived cells were obtained from 6-week old Std:ddY mouse femur of both hind limbs. The femur was harvested aseptically in a clean bench, and cut at both ends. Culture medium, consisting of DMEM (Sigma-Aldrich, Japan) supplemented with 10% fetal
Cell density
In static culture, the cell density increased with the culture period, reaching to more than double on day 7 compared to day 0 (Fig. 2(a)). The density of the amebocytes showed no marked changes during the 7-day period, while the density of the round cells greatly increased. The spindle cells increased the density on day 4, but decreased on day 7. In the case of stretch culture, an overall density was at a level of day 0 during the 7-day period (Fig. 2(b)). The density of the amebocytes
Discussion
The present study investigated mechanical regulation of differentiation capability of native bone marrow-derived cells including large amoebalike cells, round-shape cells, spindle-shape cells. When disregarding the differences in the cell shapes, there was an overall trend that the application of 10% cyclic stretch inhibited osteogenic and neurogenic differentiation, but enhanced smooth muscle differentiation. More interestingly, close examinations revealed that the responsiveness to the
Acknowledgements
This work was supported in part by KAKENHI from JSPS (Nos. 15H05860, 16K01346, 17K20102, and 18H03752) and AMED-CREST from Japan Agency for Medical Research and Development (JP19gm0810005).
Declaration of competing interest
The authors have neither financial nor personal relationships with other people or organizations that could inappropriately influence the present work.
References (39)
- et al.
In vivo measurement of human tibial strains during vigorous activity
Bone
(1996) - et al.
Matrices with compliance comparable to that of brain tissue select neuronal over glial growth in mixed cortical cultures
Biophys. J.
(2006) - et al.
Effects of cyclic stretch on proliferation of mesenchymal stem cells and their differentiation to smooth muscle cells
Biochem. Biophys. Res. Commun.
(2009) - et al.
Mechanosensitive channels and their functions in stem cell differentiation
Exp. Cell Res.
(2019) - et al.
Mechanics and mechanobiology of mesenchymal stem cell-based engineered cartilage
J. Biomech.
(2010) - et al.
Mechanical stress promotes the expression of smooth muscle-like properties in marrow stromal cells
Exp. Hematol.
(2004) - et al.
Mechanosensitive TRPM7 mediates shear stress and modulates osteogenic differentiation of mesenchymal stromal cells through Osterix pathway
Sci. Rep.
(2015) - et al.
In vitro aging of human bone marrow derived stromal cells
Mech. Ageing Dev.
(1981) - et al.
Demonstration of the presence of independent pre-osteoblastic and pre-adipocytic cell populations in bone marrow-derived mesenchymal stem cells
Bone
(2008) - et al.
Mechanical strain induces osteogenic differentiation: Cbfa1 and Ets-1 expression in stretched rat mesenchymal stem cells
Int. J. Oral Maxillofac. Surg.
(2008)
From mechanical stimulus to bone formation: a review
Med. Eng. Phys.
Cyclic tensile stress promotes osteogenic differentiation of adipose stem cells via ERK and p38 pathways
Stem Cell Res.
A rapid and efficient method for expansion of human mesenchymal stem cells
Tissue Eng.
Gene expression by marrow stromal cells in a porous collagen-glycosaminoglycan scaffold is affected by pore size and mechanical stimulation
J. Mater. Sci.–Mater. Med.
Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells
Proc. Natl. Acad. Sci.
Mechanotransduction and the functional response of bone to mechanical strain
Calcif. Tissue Int.
A collagen-glycosaminoglycan scaffold supports adult rat mesenchymal stem cell differentiation along osteogenic and chondrogenic routes
Tissue Eng.
Single-cell RNA-Seq of bone marrow-derived mesenchymal stem cells reveals unique profiles of lineage priming
PLoS ONE
Bone’s mechanostat: a 2003 update
Anat. Rec. Part A
Cited by (4)
Engineered stem cell-based strategy: A new paradigm of next-generation stem cell product in regenerative medicine
2024, Journal of Controlled ReleaseInsight into muscle stem cell regeneration and mechanobiology
2023, Stem Cell Research and TherapyHow the mechanical microenvironment of stem cell growth affects their differentiation: a review
2022, Stem Cell Research and TherapyGli1<sup>+</sup>Cells Residing in Bone Sutures Respond to Mechanical Force via IP<inf>3</inf>R to Mediate Osteogenesis
2021, Stem Cells International
- 1
Present.