Derivation of Mesenchymal Stromal Cells from Pluripotent Stem Cells through a Neural Crest Lineage using Small Molecule Compounds with Defined Media

被引：128

作者：

Fukuta, Makoto ^{[1
,2
,3
]}

Nakai, Yoshinori ^{[4
]}

Kirino, Kosuke ^{[5
]}

Nakagawa, Masato ^{[6
]}

Sekiguchi, Kazuya ^{[1
,2
,7
]}

Nagata, Sanae ^{[2
]}

Matsumoto, Yoshihisa ^{[1
,2
,3
]}

Yamamoto, Takuya ^{[6
,8
]}

Umeda, Katsutsugu ^{[9
]}

Heike, Toshio ^{[9
]}

Okumura, Naoki ^{[10
]}

Koizumi, Noriko ^{[10
]}

Sato, Takahiko ^{[4
]}

Nakahata, Tatsutoshi ^{[5
]}

Saito, Megumu ^{[5
]}

Otsuka, Takanobu ^{[3
]}

Kinoshita, Shigeru ^{[4
]}

Ueno, Morio ^{[4
]}

Ikeya, Makoto ^{[2
]}

Toguchida, Junya ^{[1
,2
,7
]}

机构：

[1] Kyoto Univ, Inst Frontier Med Sci, Dept Tissue Regenerat, Kyoto, Japan

[2] Kyoto Univ, Ctr iPS Cell Res & Applicat, Dept Cell Growth & Differentiat, Kyoto, Japan

[3] Nagoya City Univ, Grad Sch Med Sci, Dept Orthopaed Surg, Nagoya, Aichi, Japan

[4] Kyoto Prefectural Univ Med, Dept Ophthalmol, Kyoto, Japan

[5] Kyoto Univ, Dept Clin Applicat, Ctr iPS Cell Res & Applicat, Kyoto, Japan

[6] Kyoto Univ, Dept Reprogramming Sci, Ctr iPS Cell Res & Applicat, Kyoto, Japan

[7] Kyoto Univ, Grad Sch Med, Dept Orthopaed Surg, Kyoto, Japan

[8] Kyoto Univ, Inst Integrated Cell Mat Sci WPI iCeMS, Kyoto, Japan

[9] Kyoto Univ, Grad Sch Med, Dept Pediat, Kyoto, Japan

[10] Doshisha Univ, Fac Life & Med Sci, Dept Biomed Engn, Kyotanabe, Japan

来源：

PLOS ONE | 2014年 / 9卷 / 12期

关键词：

PATIENT-SPECIFIC IPSCS; PARAXIAL MESODERM; BRANCHIAL ARCHES; DIFFERENTIATION; INDUCTION; MOUSE; SPECIFICATION; MAINTENANCE; MELANOCYTES; GENERATION;

D O I：

10.1371/journal.pone.0112291

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

Neural crest cells (NCCs) are an embryonic migratory cell population with the ability to differentiate into a wide variety of cell types that contribute to the craniofacial skeleton, cornea, peripheral nervous system, and skin pigmentation. This ability suggests the promising role of NCCs as a source for cell-based therapy. Although several methods have been used to induce human NCCs (hNCCs) from human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), further modifications are required to improve the robustness, efficacy, and simplicity of these methods. Chemically defined medium (CDM) was used as the basal medium in the induction and maintenance steps. By optimizing the culture conditions, the combination of the GSK3 beta inhibitor and TGF beta inhibitor with a minimum growth factor (insulin) very efficiently induced hNCCs (70-80%) from hPSCs. The induced hNCCs expressed cranial NCC-related genes and stably proliferated in CDM supplemented with EGF and FGF2 up to at least 10 passages without changes being observed in the major gene expression profiles. Differentiation properties were confirmed for peripheral neurons, glia, melanocytes, and corneal endothelial cells. In addition, cells with differentiation characteristics similar to multipotent mesenchymal stromal cells (MSCs) were induced from hNCCs using CDM specific for human MSCs. Our simple and robust induction protocol using small molecule compounds with defined media enabled the generation of hNCCs as an intermediate material producing terminally differentiated cells for cell-based innovative medicine.

引用

页数：25

共 53 条

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Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture [J].