title: Stiffness-Controlled Thermoresponsive Hydrogels for Cell Harvesting with Sustained Mechanical Memory.
abstract: Most mechanobiological investigations focused on in situ mechanical regulation of cells on stiffness-controlled substrates with few downstream applications, as it is still challenging to harvest and expand mechanically primed cells by enzymatic digestion (e.g., trypsin) without interrupting cellular mechanical memory between passages. This study develops thermoresponsive hydrogels with controllable stiffness to generate mechanically primed cells with intact mechanical memory for augmented wound healing. No significant cellular property alteration of the fibroblasts primed on thermoresponsive hydrogels with varied stiffness has been observed through thermoresponsive harvesting. When reseeding the harvested cells for further evaluation, softer hydrogels are proven to better sustain the mechanical priming effects compared to rigid tissue culture plate, which indicates that both the stiffness-controlled substrate and thermoresponsive harvesting are required to sustain cellular mechanical memory between passages. Moreover, epigenetics analysis reveals that thermoresponsive harvesting could reduce the rearrangement and loss of chromatin proteins compared to that of trypsinization. In vivo wound healing using mechanically primed fibroblasts shows featured epithelium and sebaceous glands, which indicates augmented skin recovery compared with trypsinized fibroblasts. Thus, the thermoresponsive hydrogel-based cell harvesting system offers a powerful tool to investigate mechanobiology between cell passages and produces abundant cells with tailored mechanical priming properties for cell-based applications.
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, 100084, China.|Joint Center for Life Sciences, Tsinghua University-Peking University, Beijing, 100084, China.
authors.forename: Xingliang
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, 100084, China.|Institute of Medical Equipment, Academy of Military Medical Sciences, Tianjin, 300161, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Chemistry, School of Science, Tsinghua University, Beijing, 100084, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, 100084, China.|School of Life Science, Tsinghua University, Beijing, 100084, China.
authors.forename: Bingjie
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, 100084, China.
authors.forename: Yaozu
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Joint Center for Life Sciences, Tsinghua University-Peking University, Beijing, 100084, China.|School of Life Science, Tsinghua University, Beijing, 100084, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xie
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Beijing Tsinghua Changgung Hospital, Tsinghua University, Beijing, 102218, China.
authors.forename: Chengyu
authors.initials: C
authors.lastname: Huang
authors.affiliation: Tissue Engineering Laboratory, Department of Mechanical Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong, Special Administrative Region, China.
authors.forename: Barbara Pui
authors.initials: BP
authors.lastname: Chan
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, 100084, China.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Du
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D005347:Fibroblasts / Q000378:metabolism*; D055166:Hepatic Stellate Cells / Q000378:metabolism*; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D008819:Mice, Nude; D012867:Skin / Q000293:injuries* / Q000378:metabolism; D014945:Wound Healing
chemical_list: D020100:Hydrogels
keywords: mechanical memory; mechanobiology; stiffness-control; thermoresponsive hydrogels; wound healing