title: Non-genetic direct reprogramming and biomimetic platforms in a preliminary study for adipose-derived stem cells into corneal endothelia-like cells.
abstract: Cell fate and function can be regulated and reprogrammed by intrinsic genetic program, extrinsic factors and niche microenvironment. Direct reprogramming has shown many advantages in the field of cellular reprogramming. Here we tried the possibility to generate corneal endothelia (CE) -like cells from human adipose-derived stem cells (ADSCs) by the non-genetic direct reprogramming of recombinant cell-penetrating proteins Oct4/Klf4/Sox2 (PTD-OKS) and small molecules (purmorphamine, RG108 and other reprogramming chemical reagents), as well as biomimetic platforms of simulate microgravity (SMG) bioreactor. Co-cultured with corneal cells and decellularized corneal ECM, Reprogrammed ADSCs revealed spherical growth and positively expressing Nanog for RT-PCR analysis and CD34 for immunofluorescence staining after 7 days-treatment of both purmorphamine and PTD-OKS (P-OKS) and in SMG culture. ADSCs changed to CEC polygonal morphology from spindle shape after the sequential non-genetic direct reprogramming and biomimetic platforms. At the same time, induced cells converted to weakly express CD31, AQP-1 and ZO-1. These findings demonstrated that the treatments were able to promote the stem-cell reprogramming for human ADSCs. Our study also indicates for the first time that SMG rotary cell culture system can be used as a non-genetic means to promote direct reprogramming. Our methods of reprogramming provide an alternative strategy for engineering patient-specific multipotent cells for cellular plasticity research and future autologous CEC replacement therapy that avoids complications associated with the use of human pluripotent stem cells.
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Yonglong
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Ophthalmology Department, First Affiliated Hospital of Jinan University, Guangzhou, China; Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Ophthalmology Department, First Affiliated Hospital of Jinan University, Guangzhou, China; Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Shanyi
authors.initials: S
authors.lastname: Li
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Guo
authors.affiliation: Ophthalmology Department, First Affiliated Hospital of Jinan University, Guangzhou, China; Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, China.
authors.forename: Ruiling
authors.initials: R
authors.lastname: Lian
authors.affiliation: Bioengineering Institute of Jinan University, Guangzhou, China.
authors.forename: Rongjie
authors.initials: R
authors.lastname: Yu
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Liu
authors.affiliation: Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, China; Ophthalmology Department, First Affiliated Hospital of Jinan University, Guangzhou, China; Institute of Ophthalmology, Medical College, Jinan University, Guangzhou, China.
authors.forename: Jiansu
authors.initials: J
authors.lastname: Chen
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D000328:Adult; D000818:Animals; D032701:Biomimetics; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000502:physiology*; D004728:Endothelium, Corneal / Q000166:cytology* / Q000378:metabolism; D005260:Female; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3