Hydrogel Modulus Affects Proliferation Rate and Pluripotency of Human Mesenchymal Stem Cells Grown in Three-Dimensional Culture

自愈水凝胶 间充质干细胞 材料科学 三维细胞培养 细胞生长 细胞培养 细胞生物学 组织工程 化学 生物医学工程 生物物理学 细胞 生物 生物化学 医学 高分子化学 遗传学
作者
Revital Goldshmid,Dror Seliktar
出处
期刊:ACS Biomaterials Science & Engineering [American Chemical Society]
卷期号:3 (12): 3433-3446 被引量:37
标识
DOI:10.1021/acsbiomaterials.7b00266
摘要

Hydrogels have been used extensively with various cell types in three-dimensional (3D) culture, including with human mesenchymal stem cells (hMSCs). Here, we report on the use of poly(ethylene glycol) (PEG)-conjugated fibrinogen hydrogels to grow bone marrow-derived hMSCs in 3D culture. The initial modulus of the PEG-fibrinogen (PF) hydrogels was varied to study the influence of the stiffness on the proliferation response of the cells growing within. Shear rheology was used to quantify the changes to the initial material properties; the shear storage modulus of the PF was controlled by changing the concentration of synthetic PEG cross-linker, while keeping the fibrinogen concentration constant. Cell culture was performed during a 14-day experiment to quantify the cell proliferation response in the different modulus materials tested. The hMSCs were recovered from the hydrogels by mild enzymatic dissolution, and characterized for proliferation and cell number using cytometry. The results indicate a modulus-dependent response from the cells, and the ability to preferentially define initial hydrogel modulus that favors higher proliferation and multipotency of the hMSCs. Bioactive supplements added to the hydrogels, including exogenous fibronectin (Fn) and von willebrand factor (vWf) were used to further stimulate the proliferation response of the hMSCs in the hydrogel cultures, without altering their multipotency. These insights underscore the importance of mechanical properties in regulating cell proliferation in a 3D culture milieu. The versatility of the hydrogel culture environment and the ability to control mechanical properties for cell-fate determination further highlight the significance of a modular approach when developing materials that ultimately optimize stem cell cultures.
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