2. Academic Validation
  3. Regulation of the Keratocyte Phenotype and Cell Behavior Derived from Human Induced Pluripotent Stem Cells by Substrate Stiffness

Regulation of the Keratocyte Phenotype and Cell Behavior Derived from Human Induced Pluripotent Stem Cells by Substrate Stiffness

  • ACS Biomater Sci Eng. 2023 Jan 20. doi: 10.1021/acsbiomaterials.2c01003.
Lan Zhang 1,  Liying Liang 1,  Ting Su 1,  Yonglong Guo 1 2,  Quan Yu 3,  Deliang Zhu 4,  Zekai Cui 5,  Jun Zhang 6,  Jiansu Chen 1 5 7 8
Affiliations

Affiliations

  • 1 Ophthalmology Department, The First Affiliated Hospital of Jinan University, Guangzhou 510632, China.
  • 2 College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
  • 3 Centric Laboratory, Medical College, Jinan University, Guangzhou 510632, China.
  • 4 Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, Guangdong, PR China.
  • 5 Aier Eye Institute, Changsha 410015, Hunan Province, China.
  • 6 Key Laboratory of Optoelectronic Information and Sensing Technologies, Guangdong Higher Educational Institutes, Jinan University, Guangzhou 510632, China.
  • 7 Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou 510632, China.
  • 8 Institute of Ophthalmology, Medical College, Jinan University, Guangzhou 510632, China.
PMID: 36668685 DOI: 10.1021/acsbiomaterials.2c01003
Abstract

Substrate stiffness has been indicated as an important factor to control stem cell fate, including proliferation and differentiation. To optimize the stiffness for the differentiation process from h-iPSCs (human induced pluripotent stem cells) into h-iCSCs (human corneal stromal cells derived from h-iPSCs) and the phenotypic maintenance of h-iCSCs in vitro, h-iPSCs were cultured on matrigel-coated tissue culture plate (TCP) (106 kPa), matrigel-coated polydimethylsiloxane (PDMS) 184 (1250 kPa), and matrigel-coated PDMS 527 (4 kPa) before they were differentiated to h-iCSCs. Immunofluorescence staining, quantitative real-time polymerase chain reaction (RT-qPCR), and western blot demonstrated that the stiffer substrate TCP promoted the h-iCSCs' differentiation from h-iPSCs. On the contrary, softer PDMS 527 was more effective to maintain the phenotype of h-iCSCs cultured in vitro. Finally, we cultured h-iCSCs on PDMS 527 until P3 and seeded them on a biomimetic collagen membrane to form the single-layer and multiple-layer bioengineered corneal stroma with high transparency properties and cell survival rate. In conclusion, the study is helpful for differentiating h-iPSCs to h-iCSCs and corneal tissue engineering by manipulating stiffness mechanobiology.

Keywords

bioengineered corneal stroma; h-iPSCs; keratocytes; mechanical force; polydimethylsiloxane.

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