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Differentiation of human pluripotent stem cells to retinal pigmented epithelium in defined conditions using purified extracellular matrix proteins.
J Tissue Eng Regen Med. 2013 Aug; 7(8):642-53.JT

Abstract

A potential application of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) is the generation of retinal pigmented epithelium (RPE) to treat age-related macular degeneration (AMD), a common but incurable retinal disease. RPE cells derived from hESCs (hESC-RPEs) and iPSCs (iPSC-RPEs) express essential RPE markers and can rescue visual function in animal models. However, standard differentiation protocols yield RPE cells at low frequency, especially from iPSC lines, and the common use of Matrigel and xenogeneic feeder cells is not compatible with clinical applications. The extracellular matrix (ECM) can affect differentiation, and therefore changes in ECM composition may improve the frequency of stem cell-RPE differentiation. We selected several purified ECM proteins and substrates, based on the in vivo RPE ECM environment, and tested their ability to support iPSC-RPE differentiation and maintenance. iPSCs differentiated on nearly all tested substrates developed pigmented regions, with Matrigel and mouse laminin-111 supporting the highest pigmentation frequencies. Although iPSC-RPEs cultured on the majority of the tested substrates expressed key RPE genes, only six substrates supported development of confluent monolayers with normal RPE morphology, including Matrigel and mouse laminin-111. iPSCs differentiated on mouse laminin-111 produced iPSC-RPEs expressing RPE proteins, and hESCs differentiated on mouse laminin-111 resulted in high yields of functional hESC-RPEs. This identification of key ECM proteins may assist with future scaffold designs and provide peptide sequences for use in synthetic, xeno-free, GMP-compliant generation of RPE from human pluripotent stem cells relevant to clinical translation.

Authors+Show Affiliations

Center for Stem Cell Biology and Engineering, University of California, Santa Barbara, CA 93106, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

22514096

Citation

Rowland, Teisha J., et al. "Differentiation of Human Pluripotent Stem Cells to Retinal Pigmented Epithelium in Defined Conditions Using Purified Extracellular Matrix Proteins." Journal of Tissue Engineering and Regenerative Medicine, vol. 7, no. 8, 2013, pp. 642-53.
Rowland TJ, Blaschke AJ, Buchholz DE, et al. Differentiation of human pluripotent stem cells to retinal pigmented epithelium in defined conditions using purified extracellular matrix proteins. J Tissue Eng Regen Med. 2013;7(8):642-53.
Rowland, T. J., Blaschke, A. J., Buchholz, D. E., Hikita, S. T., Johnson, L. V., & Clegg, D. O. (2013). Differentiation of human pluripotent stem cells to retinal pigmented epithelium in defined conditions using purified extracellular matrix proteins. Journal of Tissue Engineering and Regenerative Medicine, 7(8), 642-53. https://doi.org/10.1002/term.1458
Rowland TJ, et al. Differentiation of Human Pluripotent Stem Cells to Retinal Pigmented Epithelium in Defined Conditions Using Purified Extracellular Matrix Proteins. J Tissue Eng Regen Med. 2013;7(8):642-53. PubMed PMID: 22514096.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Differentiation of human pluripotent stem cells to retinal pigmented epithelium in defined conditions using purified extracellular matrix proteins. AU - Rowland,Teisha J, AU - Blaschke,Alison J, AU - Buchholz,David E, AU - Hikita,Sherry T, AU - Johnson,Lincoln V, AU - Clegg,Dennis O, Y1 - 2012/04/18/ PY - 2011/06/06/received PY - 2011/10/05/revised PY - 2011/11/24/accepted PY - 2012/4/20/entrez PY - 2012/4/20/pubmed PY - 2013/11/2/medline KW - age-related macular degeneration KW - extracellular matrix KW - human embryonic stem cells KW - induced pluripotent stem cells KW - integrins KW - laminin KW - pluripotent stem cells KW - retinal pigmented epithelium SP - 642 EP - 53 JF - Journal of tissue engineering and regenerative medicine JO - J Tissue Eng Regen Med VL - 7 IS - 8 N2 - A potential application of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) is the generation of retinal pigmented epithelium (RPE) to treat age-related macular degeneration (AMD), a common but incurable retinal disease. RPE cells derived from hESCs (hESC-RPEs) and iPSCs (iPSC-RPEs) express essential RPE markers and can rescue visual function in animal models. However, standard differentiation protocols yield RPE cells at low frequency, especially from iPSC lines, and the common use of Matrigel and xenogeneic feeder cells is not compatible with clinical applications. The extracellular matrix (ECM) can affect differentiation, and therefore changes in ECM composition may improve the frequency of stem cell-RPE differentiation. We selected several purified ECM proteins and substrates, based on the in vivo RPE ECM environment, and tested their ability to support iPSC-RPE differentiation and maintenance. iPSCs differentiated on nearly all tested substrates developed pigmented regions, with Matrigel and mouse laminin-111 supporting the highest pigmentation frequencies. Although iPSC-RPEs cultured on the majority of the tested substrates expressed key RPE genes, only six substrates supported development of confluent monolayers with normal RPE morphology, including Matrigel and mouse laminin-111. iPSCs differentiated on mouse laminin-111 produced iPSC-RPEs expressing RPE proteins, and hESCs differentiated on mouse laminin-111 resulted in high yields of functional hESC-RPEs. This identification of key ECM proteins may assist with future scaffold designs and provide peptide sequences for use in synthetic, xeno-free, GMP-compliant generation of RPE from human pluripotent stem cells relevant to clinical translation. SN - 1932-7005 UR - https://www.unboundmedicine.com/medline/citation/22514096/Differentiation_of_human_pluripotent_stem_cells_to_retinal_pigmented_epithelium_in_defined_conditions_using_purified_extracellular_matrix_proteins_ L2 - https://doi.org/10.1002/term.1458 DB - PRIME DP - Unbound Medicine ER -