Suppr超能文献

在定义的培养条件下从人诱导多能干细胞生成角质形成细胞。

Generation of Keratinocytes from Human Induced Pluripotent Stem Cells Under Defined Culture Conditions.

机构信息

Center for Regenerative Medicine and Skeletal Development, Department of Reconstructive Sciences, University of Connecticut Health, Farmington, Connecticut, USA.

Department of Oral Health and Diagnostic Sciences, School of Dental Medicine, University of Connecticut Health, Farmington, Connecticut, USA.

出版信息

Cell Reprogram. 2021 Feb;23(1):1-13. doi: 10.1089/cell.2020.0046. Epub 2020 Dec 29.

Abstract

Differentiation of keratinocytes from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) has become an important tool for wound healing research and for studying skin diseases in instances where patient cells are not available. Several keratinocyte differentiation protocols using hiPSC colony fragments or embryoid bodies have been published with some requiring prolonged time for differentiation or extended use of reagent cocktails. In this study, we present a simplified method to efficiently generate large numbers of uniformly differentiated keratinocytes in less than 4 weeks from singularized hiPSCs with differentiation factors, retinoic acid and bone morphogenetic protein 4 (BMP4). Low seeding density of singularized iPSCs results in keratinocyte cultures with minimum cell death during differentiation and up to 96% homogeneity for keratin 14-positive cells and low percentage of keratinocyte maturation markers, comparable to early passage primary keratinocytes. hiPSC-derived keratinocytes remain in a proliferative state, can be maintained for prolonged periods of time, and can be terminally differentiated under high calcium conditions in the same way as primary human keratinocytes. Moreover, coculturing hiPSC-derived fibroblasts and keratinocytes consistently formed organotypic 3D skin equivalents. Therefore, keratinocytes generated by this method are a viable source of cells for downstream applications.

摘要

从人类胚胎干细胞(hESC)和人类诱导多能干细胞(hiPSC)分化角质形成细胞已成为伤口愈合研究的重要工具,并且在无法获得患者细胞的情况下,也可用于研究皮肤疾病。已经发表了几种使用 hiPSC 集落片段或类胚体的角质形成细胞分化方案,其中一些需要长时间分化或延长使用试剂混合物。在这项研究中,我们提出了一种简化的方法,可在不到 4 周的时间内,从单个 hiPSC 与分化因子、维甲酸和骨形态发生蛋白 4(BMP4)高效产生大量均匀分化的角质形成细胞。单个 hiPSC 的低接种密度可导致分化过程中细胞死亡最小的角质形成细胞培养物,角蛋白 14 阳性细胞的均匀性高达 96%,且角质形成细胞成熟标志物的百分比低,与早期传代的原代角质形成细胞相当。hiPSC 衍生的角质形成细胞保持增殖状态,可长时间维持,并可在高钙条件下以与原代人角质形成细胞相同的方式进行终末分化。此外,hiPSC 衍生的成纤维细胞和角质形成细胞的共培养始终形成器官型 3D 皮肤等效物。因此,通过该方法产生的角质形成细胞是下游应用的可行细胞来源。

相似文献

1
Generation of Keratinocytes from Human Induced Pluripotent Stem Cells Under Defined Culture Conditions.
Cell Reprogram. 2021 Feb;23(1):1-13. doi: 10.1089/cell.2020.0046. Epub 2020 Dec 29.
2
Deriving Keratinocyte Progenitor Cells and Keratinocytes from Human-Induced Pluripotent Stem Cells.
Curr Protoc Stem Cell Biol. 2020 Sep;54(1):e119. doi: 10.1002/cpsc.119.
4
Generation of functional multipotent keratinocytes from mouse induced pluripotent stem cells.
Methods Mol Biol. 2013;961:337-50. doi: 10.1007/978-1-62703-227-8_22.
5
Directed differentiation of human embryonic stem cells into keratinocyte progenitors in vitro: an attempt with promise of clinical use.
In Vitro Cell Dev Biol Anim. 2016 Sep;52(8):885-93. doi: 10.1007/s11626-016-0024-2. Epub 2016 Aug 5.
6
Self-assembly of differentiated progenitor cells facilitates spheroid human skin organoid formation and planar skin regeneration.
Theranostics. 2021 Jul 25;11(17):8430-8447. doi: 10.7150/thno.59661. eCollection 2021.
7
Melanin Transfer in Human 3D Skin Equivalents Generated Exclusively from Induced Pluripotent Stem Cells.
PLoS One. 2015 Aug 26;10(8):e0136713. doi: 10.1371/journal.pone.0136713. eCollection 2015.
8
Differentiation of human induced pluripotent stem cells into a keratinocyte lineage.
Methods Mol Biol. 2014;1195:1-12. doi: 10.1007/7651_2013_64.

引用本文的文献

2
Organoids in skin wound healing.
Burns Trauma. 2025 Jan 3;13:tkae077. doi: 10.1093/burnst/tkae077. eCollection 2025.
3
Advances in Microengineered Platforms for Skin Research.
JID Innov. 2024 Sep 18;5(1):100315. doi: 10.1016/j.xjidi.2024.100315. eCollection 2025 Jan.
5
Human-Induced Pluripotent Stem Cells in Plastic and Reconstructive Surgery.
Int J Mol Sci. 2024 Feb 3;25(3):1863. doi: 10.3390/ijms25031863.
6
A Systematic Review of Stem Cell Differentiation into Keratinocytes for Regenerative Applications.
Stem Cell Rev Rep. 2024 Jan;20(1):362-393. doi: 10.1007/s12015-023-10636-9. Epub 2023 Nov 3.
7
iPSC-based approach for human hair follicle regeneration.
Front Cell Dev Biol. 2023 May 30;11:1149050. doi: 10.3389/fcell.2023.1149050. eCollection 2023.
8
Direct Lineage Reprogramming for Induced Keratinocyte Stem Cells: A Potential Approach for Skin Repair.
Stem Cells Transl Med. 2023 May 15;12(5):245-257. doi: 10.1093/stcltm/szad019.
9
Differentiation of pluripotent stem cells for modeling human skin development and potential applications.
Front Cell Dev Biol. 2022 Nov 23;10:1030339. doi: 10.3389/fcell.2022.1030339. eCollection 2022.

本文引用的文献

1
CRISPR/Cas9-based targeted genome editing for correction of recessive dystrophic epidermolysis bullosa using iPS cells.
Proc Natl Acad Sci U S A. 2019 Dec 26;116(52):26846-26852. doi: 10.1073/pnas.1907081116. Epub 2019 Dec 9.
2
Induced pluripotent stem cell-derived melanocyte precursor cells undergoing differentiation into melanocytes.
Pigment Cell Melanoma Res. 2019 Sep;32(5):623-633. doi: 10.1111/pcmr.12779. Epub 2019 Mar 25.
3
Epithelial barrier repair and prevention of allergy.
J Clin Invest. 2019 Apr 1;129(4):1463-1474. doi: 10.1172/JCI124608. Epub 2019 Feb 18.
4
Prolonging culture of primary human keratinocytes isolated from suction blisters with the Rho kinase inhibitor Y-27632.
PLoS One. 2018 Sep 12;13(9):e0198862. doi: 10.1371/journal.pone.0198862. eCollection 2018.
5
In vivo reprogramming of wound-resident cells generates skin epithelial tissue.
Nature. 2018 Sep;561(7722):243-247. doi: 10.1038/s41586-018-0477-4. Epub 2018 Sep 5.
7
3D skin models for 3R research: The potential of 3D reconstructed skin models to study skin barrier function.
Exp Dermatol. 2018 May;27(5):501-511. doi: 10.1111/exd.13531. Epub 2018 Apr 6.
9
Regeneration of the entire human epidermis using transgenic stem cells.
Nature. 2017 Nov 16;551(7680):327-332. doi: 10.1038/nature24487. Epub 2017 Nov 8.
10
Craniometaphyseal Dysplasia Mutations in ANKH Negatively Affect Human Induced Pluripotent Stem Cell Differentiation into Osteoclasts.
Stem Cell Reports. 2017 Nov 14;9(5):1369-1376. doi: 10.1016/j.stemcr.2017.09.016. Epub 2017 Oct 19.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验