Xeno-free and feeder-free culture and differentiation of human embryonic stem cells on recombinant vitronectin-grafted hydrogels

Chen LH 1 , Sung TC 1 , Lee HH 2 , Higuchi A 3 , Su HC 1 , Lin KJ 1 Show all authors , Huang YR 1 , Ling QD 4 , Kumar SS 5 , Alarfaj AA 6 , Munusamy MA 6 , Nasu M 7 , Chen DC 8 , Hsu ST 9 , Chang Y 10 , Lee KF 11 , Wang HC 12 , Umezawa A 7

Affiliations 

  • 1 Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001, Taiwan. higuchi@ncu.edu.tw
  • 2 Department of Surgery, Hsinchu Cathay General Hospital, No. 678, Sec 2, Zhonghua Rd., Hsinchu, 30060, Taiwan and Graduate Institute of Translational and Interdisciplinary Medicine, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001, Taiwan
  • 3 Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001, Taiwan. higuchi@ncu.edu.tw and Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia and Department of Reproduction, National Center for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan and Department of Chemical Engineering, R&D Center for Membrane Technology, Chung Yuan Christian University, 200, Chung-Bei Rd., Chungli, Taoyuan, 320, Taiwan
  • 4 Cathay Medical Research Institute, Cathay General Hospital, No. 32, Ln 160, Jian-Cheng Road, Hsi-Chi City, Taipei, 221, Taiwan
  • 5 Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, 43400 Serdang, Slangor, Malaysia
  • 6 Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia
  • 7 Department of Reproduction, National Center for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan
  • 8 Department of Obstetrics and Gynecology, Taiwan Landseed Hospital, 77, Kuangtai Road, Pingjen City, Taoyuan 32405, Taiwan
  • 9 Department of Internal Medicine, Taiwan Landseed Hospital, 77, Kuangtai Road, Pingjen City, Taoyuan 32405, Taiwan
  • 10 Department of Chemical Engineering, R&D Center for Membrane Technology, Chung Yuan Christian University, 200, Chung-Bei Rd., Chungli, Taoyuan, 320, Taiwan
  • 11 Precision Medical laboratory, Lee's OB/GYN Clinic, No. 9, Ln. 31, Sec. 2, Jinshan S. Rd., Da'an Dist., Taipei 106, Taiwan
  • 12 Department of Obstetrics and Gynecology, Hungchi Women & Children's Hospital, Taoyuan 320, Taiwan
Biomater Sci, 2019 Aug 14.
PMID: 31411209 DOI: 10.1039/c9bm00418a

Abstract

Recombinant vitronectin-grafted hydrogels were developed by adjusting surface charge of the hydrogels with grafting of poly-l-lysine for optimal culture of human embryonic stem cells (hESCs) under xeno- and feeder-free culture conditions, with elasticity regulated by crosslinking time (10-30 kPa), in contrast to conventional recombinant vitronectin coating dishes, which have a fixed stiff surface (3 GPa). hESCs proliferated on the hydrogels for over 10 passages and differentiated into the cells derived from three germ layers indicating the maintenance of pluripotency. hESCs on the hydrogels differentiated into cardiomyocytes under xeno-free culture conditions with much higher efficiency (80% of cTnT+ cells) than those on conventional recombinant vitronectin or Matrigel-coating dishes just only after 12 days of induction. It is important to have an optimal design of cell culture biomaterials where biological cues (recombinant vitronectin) and physical cues (optimal elasticity) are combined for high differentiation of hESCs into specific cell lineages, such as cardiomyocytes, under xeno-free and feeder-free culture conditions.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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