Engineered cell-laden thermosensitive poly(N-isopropylacrylamide)-immobilized gelatin microspheres as 3D cell carriers for regenerative medicine

I-Hsuan Yang; Che-Yung Kuan; Zhi-Yu Chen; Chi-Han Li; Chih-Ying Chi; Yu-Ying Lin; Ya-Jyun Liang; Wei-Ting Kuo; Yi-An Li; Feng-Huei Lin

doi:10.1016/j.mtbio.2022.100266

Engineered cell-laden thermosensitive poly(N-isopropylacrylamide)-immobilized gelatin microspheres as 3D cell carriers for regenerative medicine

Mater Today Bio. 2022 Apr 19:15:100266. doi: 10.1016/j.mtbio.2022.100266. eCollection 2022 Jun.

Authors

I-Hsuan Yang^{1

2}, Che-Yung Kuan^{1

2}, Zhi-Yu Chen^{1

2}, Chi-Han Li^{2

3}, Chih-Ying Chi⁴, Yu-Ying Lin^{2

3}, Ya-Jyun Liang¹, Wei-Ting Kuo¹, Yi-An Li¹, Feng-Huei Lin^{1

2

3}

Affiliations

¹ Department of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, No. 49, Fanglan Rd, Taipei, 10672, Taiwan.
² Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, No. 35, Keyan Road, Zhunan, Miaoli County, 35053, Taiwan.
³ PhD Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, Taichung, Taiwan.
⁴ Biomaterials Translational Research Center, China Medical University Hospital, Taiwan.

Abstract

Several studies have focused on using cell carriers to solve the problem of mesenchymal stem cell expansion on regenerative medicine. However, the disadvantages of using prolonged enzymatic treatment and low cell harvest efficiency still trouble researchers. In this study, PNIPAAm-immobilized gelatin microspheres (abbreviated as GNMS) were synthesized using a simple power-driven flow-focusing microinjection system. The developed thermosensitive GNMS can allow easier harvesting of cells from the microspheres, requiring only 10 min of low-temperature treatment and 5 min of trypsin treatment. The developed GNMS was characterized by Fourier-transform infrared spectroscopy, optical microscopy, and scanning electron microscopy. Further, live/dead staining, F-actin staining, and PrestoBlue cell viability assays were used to evaluate cytotoxicity, cell morphology, cell proliferation, and harvest efficiency. The gene expression of stem cell markers was determined by real-time quantitative PCR (Q-PCR) analysis to investigate the stemness and phenotypic changes in Wharton's jelly-derived mesenchymal stem cells. The results showed that the engineered cell-laden thermosensitive GNMS could significantly increase the cell harvest rate with over 99% cell survival rate and no change in the cell phenotype. Thus, the described strategy GNMS could be the suitable 3D cell carriers in the therapeutic application and opens new avenues for regenerative medicine.

Keywords: Cell expansion; Gelatin-PNIPAAm microspheres; Regenerative medicine; Stem cell therapy; Thermosensitive cell carriers.