Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels

Yufan Liu; Zhao Li; Jianjun Li; Siming Yang; Yijie Zhang; Bin Yao; Wei Song; Xiaobing Fu; Sha Huang

doi:10.1093/burnst/tkaa029

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Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels

更新时间：2022-06-09

- Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels
- Burns & Trauma Vol. 8, Issue 1, (2020)
- 作者机构：
  
  1. Research Center for Tissue Repair and Regeneration affiliated to the Medical Innovation Research Department
  2. PLA General Hospital and PLA Medical College
  3. PLA Key Laboratory of Tissue Repair and Regenerative Medicine and Beijing Key Research Laboratory of Skin Injury
- 作者简介：
- 基金信息：
- DOI：10.1093/burnst/tkaa029
  CLC：
- Published：2020，
  
  Published Online：27 July 2020，
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Yufan Liu, Zhao Li, Jianjun Li, Siming Yang, Yijie Zhang, Bin Yao, Wei Song, Xiaobing Fu, Sha Huang, Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels, Burns & Trauma, Volume 8, 2020, tkaa029, https://doi.org/10.1093/burnst/tkaa029
DOI：

Yufan Liu, Zhao Li, Jianjun Li, Siming Yang, Yijie Zhang, Bin Yao, Wei Song, Xiaobing Fu, Sha Huang, Stiffness-mediated mesenchymal stem cell fate decision in 3D-bioprinted hydrogels, Burns & Trauma, Volume 8, 2020, tkaa029, https://doi.org/10.1093/burnst/tkaa029 DOI：

摘要

Abstract

BackgroundHydrogels with tuneable mechanical properties are an attractive material platform for 3D bioprinting. Thus far

numerous studies have confirmed that the biophysical cues of hydrogels

such as stiffness

are known to have a profound impact on mesenchymal stem cell (MSC) differentiation; however

their differentiation potential within 3D-bioprinted hydrogels is not completely understood. Here

we propose a protocol for the exploration of how the stiffness of alginate–gelatin (Alg-Gel) composite hydrogels (the widely used bioink) affects the differentiation of MSCs in the presence or absence of differentiation inducing factors.MethodsTwo types of Alg-Gel composite hydrogels (Young’s modulus: 50 kPa vs. 225 kPa) were bioprinted independently of porosity. Then

stiffness-induced biases towards adipogenic and osteogenic differentiation of the embedded MSCs were analysed by co-staining with alkaline phosphatase (ALP) and oil red O. The expression of specific markers at the gene level was detected after a 3-day culture.ResultsConfocal microscopy indicated that all tested hydrogels supported MSC growth and viability during the culture period. Higher expression of adipogenic and osteogenic markers (ALP and lipoprotein lipase (LPL)) in stiffer 3D-bioprinted matrices demonstrated a more significant response of MSCs to stiffer hydrogels with respect to differentiation

which was more robust in differentiation-inducing medium. However

the LPL expression in stiffer 3D-bioprinted constructs was reduced at day 3 regardless of the presence of differentiation-inducing factors. Although MSCs embedded in softer hydrogels to some extent proceeded toward adipogenic and osteogenic lineages within a few days

their differentiation seemed to be slower and more limited. Interestingly

the hydrogel itself (without differentiation-inducing factors) exhibited a slight effect on whether MSCs differentiated towards an adipogenic or an osteogenic fate. Considering that the mechano-regulated protein Yes-associated protein (YAP) is involved in MSC fate decisions

we further found that inhibition of YAP significantly downregulated the expression of ALP and LPL in MSCs in stiffer constructs regardless of the induced growth factors present.ConclusionsThese results demonstrate that the differentiation of MSCs in 3D-bioprinted matrices is dependent on hydrogel stiffness

which emphasizes the importance of biophysical cues as a determinant of cellular behaviour.

关键词

Keywords

Mesenchymal stem cellsStiffnessAdipogenesisOsteogenesisDifferentiation3D-bioprinting

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