Cell migration in three-dimensional environment is extremely important for tissue regeneration and other biological processes.In this work,a model system was developed to study how endothelial cells(ECs)migrate into p...Cell migration in three-dimensional environment is extremely important for tissue regeneration and other biological processes.In this work,a model system was developed to study how endothelial cells(ECs)migrate into photo-responsive hydrogels under the presence of pro-inflammatory macrophages.The hydrogel was synthesized from hyaluronic acid grafted with coumarin and methacrylate moieties by both carbon–carbon covalent linking and coumarin dimerization under UV irradiation at 365 nm.The structure of the hydrogel was conveniently modulated by UV irradiation at 254nm to decompose the coumarin dimers,leading to the significant decrease of modulus and increase of swelling ratio and mesh size.Under the presence of M1 macrophages,ECs were induced to migrate into the hydrogels with a different degree.A significant larger net displacement of ECs was found in the softer hydrogel obtained by irradiation with UV at 254nm than in the stiffer original one at day 7.展开更多
Cell migration plays a pivotal role in many pathological and physiological processes.So far,most of the studies have been focused on 2-dimensional cell adhesion and migration.Herein,the migration behaviors of cell sph...Cell migration plays a pivotal role in many pathological and physiological processes.So far,most of the studies have been focused on 2-dimensional cell adhesion and migration.Herein,the migration behaviors of cell spheroids in 3D hydrogels obtained by polymerization of methacrylated hyaluronic acid(HA-MA)and fibrinogen(Fg)with different ratios were studied.The Fg could be released to the medium gradually along with time prolongation,achieving the dynamic change of hydrogel structures and properties.Three types of cell spheroids,i.e.,endothelial cell(EC),smooth muscle cell(SMC),and EC-SMC spheroids,were prepared with 10,000 cells in each,whose diameters were about 343,108,and 224μm,respectively.The composite hydrogels with an intermediate ratio of Fg allowed the fastest 3D migration of cell spheroids.The ECs-SMCs migrated longest up to 3200μm at day 14,whereas the SMC spheroids migrated slowest with a distance of only~400μm at the same period of time.The addition of free RGD or anti-CD44 could significantly reduce the migration distance,revealing that the cell-substrate interactions take the major roles and the migration is mesenchymal dependent.Moreover,addition of anti-N-cadherin and MMP inhibitors also slowed down the migration rate,demonstrating that the degradation of hydrogels and cell-cell interactions are also largely involved in the cell migration.RT-PCR measurement showed that expression of genes related to cell adhesion and antiapoptosis,and angiogenesis was all upregulated in the EC-SMC spheroids than single EC or SMC spheroids,suggesting that the use of composite cell spheroids is more promising to promote cell-substrate interactions and maintenance of cell functions.展开更多
基金Natural Science Foundation of China(51873188,21434006).
文摘Cell migration in three-dimensional environment is extremely important for tissue regeneration and other biological processes.In this work,a model system was developed to study how endothelial cells(ECs)migrate into photo-responsive hydrogels under the presence of pro-inflammatory macrophages.The hydrogel was synthesized from hyaluronic acid grafted with coumarin and methacrylate moieties by both carbon–carbon covalent linking and coumarin dimerization under UV irradiation at 365 nm.The structure of the hydrogel was conveniently modulated by UV irradiation at 254nm to decompose the coumarin dimers,leading to the significant decrease of modulus and increase of swelling ratio and mesh size.Under the presence of M1 macrophages,ECs were induced to migrate into the hydrogels with a different degree.A significant larger net displacement of ECs was found in the softer hydrogel obtained by irradiation with UV at 254nm than in the stiffer original one at day 7.
基金This study is financially supported by the Natural Science Foundation of China(51873188,21434006)the National Key Research and Development Program of China(2016YFC1100403).
文摘Cell migration plays a pivotal role in many pathological and physiological processes.So far,most of the studies have been focused on 2-dimensional cell adhesion and migration.Herein,the migration behaviors of cell spheroids in 3D hydrogels obtained by polymerization of methacrylated hyaluronic acid(HA-MA)and fibrinogen(Fg)with different ratios were studied.The Fg could be released to the medium gradually along with time prolongation,achieving the dynamic change of hydrogel structures and properties.Three types of cell spheroids,i.e.,endothelial cell(EC),smooth muscle cell(SMC),and EC-SMC spheroids,were prepared with 10,000 cells in each,whose diameters were about 343,108,and 224μm,respectively.The composite hydrogels with an intermediate ratio of Fg allowed the fastest 3D migration of cell spheroids.The ECs-SMCs migrated longest up to 3200μm at day 14,whereas the SMC spheroids migrated slowest with a distance of only~400μm at the same period of time.The addition of free RGD or anti-CD44 could significantly reduce the migration distance,revealing that the cell-substrate interactions take the major roles and the migration is mesenchymal dependent.Moreover,addition of anti-N-cadherin and MMP inhibitors also slowed down the migration rate,demonstrating that the degradation of hydrogels and cell-cell interactions are also largely involved in the cell migration.RT-PCR measurement showed that expression of genes related to cell adhesion and antiapoptosis,and angiogenesis was all upregulated in the EC-SMC spheroids than single EC or SMC spheroids,suggesting that the use of composite cell spheroids is more promising to promote cell-substrate interactions and maintenance of cell functions.
