Directing three-dimensional multicellular morphogenesis by self-organization of vascular mesenchymal cells in hyaluronic acid hydrogels

Xiaolu Zhu,Chih-Ming Ho,Ting-Hsuan Chen,Siyan Dong,Peng Fei,Tatiana Segura,Shiva Gojgini

doi:10.1186/s13036-017-0055-6

Abstract

BackgroundPhysical scaffolds are useful for supporting cells to form three-dimensional (3D) tissue. However, it is non-trivial to develop a scheme that can robustly guide cells to self-organize into a tissue with the desired 3D spatial structures. To achieve this goal, the rational regulation of cellular self-organization in 3D extracellular matrix (ECM) such as hydrogel is needed.ResultsIn this study, we integrated the Turing reaction–diffusion mechanism with the self-organization process of cells and produced multicellular 3D structures with the desired configurations in a rational manner. By optimizing the components of the hydrogel and applying exogenous morphogens, a variety of multicellular 3D architectures composed of multipotent vascular mesenchymal cells (VMCs) were formed inside hyaluronic acid (HA) hydrogels. These 3D architectures could mimic the features of trabecular bones and multicellular nodules. Based on the Turing reaction–diffusion instability of morphogens and cells, a theoretical model was proposed to predict the variations observed in 3D multicellular structures in response to exogenous factors. It enabled the feasibility to obtain diverse types of 3D multicellular structures by addition of Noggin and/or BMP2.ConclusionsThe morphological consistency between the simulation prediction and experimental results probably revealed a Turing-type mechanism underlying the 3D self-organization of VMCs in HA hydrogels. Our study has provided new ways to create a variety of self-organized 3D multicellular architectures for regenerating biomaterial and tissues in a Turing mechanism-based approach.

Highlights

Physical scaffolds are useful for supporting cells to form three-dimensional (3D) tissue
The final product hyaluronic acid (HA)-AC was analyzed with 1H NMR (D2O) and the degree of acrylation (16%) was determined by dividing the multiplet peak at δ = 6.2, which corresponds to cis and trans acrylate hydrogens, by the singlet peak at δ = 1.6, which corresponds to acetylmethyl protons in HA
The network of cells became more pronounced between day 4 and 8 (Additional file 2: Figure S3b–d). 3D multicellular networks that consisted of a large number of branches and sub-branches, formed by local multicellular aggregation at day 7 (Fig. 1d)

Summary

Introduction

Physical scaffolds are useful for supporting cells to form three-dimensional (3D) tissue. Engineered three-dimensional (3D) hydrogel matrices [8, 21] provide a physiological context that closely mimics the natural ECM; events such as intercellular communication, cell-matrix interactions and cellular self-assembly can be recapitulated in defined 3D matrices [22, 23] These 3D culture strategies have produced some multicellular structures, including spheroids consisting of tumor or stem cells [24], lumen formed by glandular cells [24], bronchioalveolar-like branching networks [25], optic-cup-like structures [4], tubular structures [26, 27], and even liver organ buds [19]. These attempts are typically more laborious than those involving self-organization of cells in a simple 3D hydrogel system without external assistance

Methods

Results

Conclusion