Reemergence of undifferentiated cells from transplants of human induced pluripotent stem cells is a possible potential risk factor of tumorigenic differentiation

Kumazaki, Tatsuo; Takahashi, Tomoko; Matsuo, Taira; Kamada, Mizuna; Mitsui, Youji

doi:10.1002/cbi3.10012

Cited by 4 publications

(9 citation statements)

References 28 publications

(36 reference statements)

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“…The formed tumor was fixed with 4 % paraformaldehyde solution to examine histopathology by staining with hematoxylin and eosin at Sapporo General Pathology Laboratory Co., Ltd. (Sapporo, Japan). Another part of the fresh tumor was subjected for cell isolation as described before [ 10 ]. We cultured some of the isolated cells (K12te, K17te) at an extremely low density for 2 weeks and then picked the colonies formed from single cells into dishes as clones and expanded them for further examination.…”

Section: Methodsmentioning

confidence: 99%

“…We purified the total RNA to perform PCR with an appropriate primer set and Primestar DNA polymerase (Takara Bio, Japan) as described previously [ 10 ]. The primer sequences used are given in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

“…Slides were observed with Axiovert 200 M fluorescence microscope system (Carl Zeiss, Germany) after staining. Details are described previously [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

“…However, unknown mechanisms seem to exist in emergence of transformed cells from iPSC derivatives. Our recent study [ 10 ] on re-emergence of undifferentiated cells from hiPSC-derivatives prompted us to examine risks of transformation and de-differentiation of the differentiated cells. Further, a series of studies on cellular aging [ 11 ], immortalization [ 12 ], de-differentiation [ 10 ], and reprogramming [ 13 ] of human fibroblast TIG-1, urged us to isolate transformed cells from TIG-1 iPSC-teratomas and to disclose novel tumor risks of hiPSC-derivatives.…”

Section: Introductionmentioning

confidence: 99%

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Reversible transformation and de-differentiation of human cells derived from induced pluripotent stem cell teratomas

et al. 2015

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We first aimed to generate transformed cell lines from a human induced pluripotent stem cell (hiPSC)-teratoma, and then examined the tumorigenic risks of the differentiated cells from hiPSC explant, because hiPSC-derivatives give rise to tumors in immune-deficient mice when transplanted. The colonies isolated from sparse cultures of hiPSC-teratoma cells expressed NANOG and OCT3/4 strongly, and telomerase reverse transcriptase (TERT) weakly. However, soft agar assay demonstrated that only one of them generated colonies in the gel, though hiPSCs, hTERT-transfected immortal cells, and its oncogene-transfected cells did not form any colonies. Furthermore, none of colonies isolated from the soft agar gel on primary culture (passage 0) of teratoma cells, expressed NANOG and OCT3/4 in the expanded cultures. The second soft agar assay on the colony-derived cells was unexpectedly negative. The cumulative growth curve, telomere shortening, and senescence-associated β-galactosidase (SA β-gal) staining confirmed the mortality of these cells, suggesting their reversible transformation. By using medium for embryonic stem cell (ESC medium) after MCDB 131 (MCDB) medium, the differentiated culture cells derived from hiPSC-teratoma converted into the cells expressing undifferentiated marker proteins, which lost afterwords even in ESC medium with feeder SNL76/7. The reversibility of transformation and de-differentiation suggest that tumorigenic risks of differentiated cells arise when they are exposed to suitable niches in vivo. Thus, removal of only the undifferentiated cells from iPSC-derivatives before transplantation does not solve the problem. Elucidation of mechanisms of reversibility and control of epigenetic changes is discussed as a safety bottleneck for hiPSC therapy.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Slides were observed with Axiovert 200 M fluorescence microscope system (Carl Zeiss, Germany) after staining. Details are described previously [ 10 ].…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reversible transformation and de-differentiation of human cells derived from induced pluripotent stem cell teratomas

et al. 2015

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“…Functional cartilage is spontaneously produced from human iPSCs in teratomas (Kumazaki et al, 2014;Takahashi et al, 2007b;Yu et al, 2007), making these cells possible candidates for cartilage regenerative medicine applications and a tool for studying disease mechanisms and new treatments.…”

Section: Introductionmentioning

confidence: 99%

Induced pluripotent stem cells for cartilage repair: current status and future perspectives

Castro-Viñuelas¹,

Sanjurjo‐Rodríguez²,

Piñeiro-Ramil³

et al. 2018

eCM

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The establishment of cartilage regenerative medicine is an important clinical issue, but the search for cell sources able to restore cartilage integrity proves to be challenging. Human mesenchymal stromal cells (MSCs) are prone to form epiphyseal or hypertrophic cartilage and have an age-related limited proliferation. On the other hand, it is difficult to obtain functional chondrocytes from human embryonic stem cells (ESCs). Moreover, the ethical issues associated with human ESCs are an additional disadvantage of using such cells. Since their discovery in 2006, induced pluripotent stems cells (iPSCs) have opened many gateways to regenerative medicine research, especially in cartilage tissue engineering therapies. iPSCs have the capacity to overcome limitations associated with current cell sources since large numbers of autologous cells can be derived from small starting populations. Moreover, problems associated with epiphyseal or hypertrophiccartilage formation can be overcome using iPSCs. iPSCs emerge as a promising cell source for treating cartilage defects and have the potential to be used in the clinical field. For this purpose, robust protocols to induce chondrogenesis, both in vitro an in vivo, are required. This review summarises the recent progress in iPSC technology and its applications for cartilage repair.

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Bio-3D printing iPSC-derived human chondrocytes for articular cartilage regeneration

et al. 2021

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Osteoarthritis is a leading cause of pain and joint immobility, the incidence of which is increasing worldwide. Currently, total joint replacement is the only treatment for end-stage disease. Scaffold-based tissue engineering is a promising alternative approach for joint repair but is subject to limitations such as poor cytocompatibility and degradation-associated toxicity. To overcome these limitations, a completely scaffold-free Kenzan method for bio-3D printing was used to fabricate cartilage constructs feasible for repairing large chondral defects. Human induced pluripotent stem cell (iPSC)-derived neural crest cells with high potential to undergo chondrogenesis through mesenchymal stem cell differentiation were used to fabricate the cartilage. Unified, self-sufficient, and functional cartilaginous constructs up to 6 cm2 in size were assembled by optimizing fabrication time during chondrogenic induction. Maturation for 3 weeks facilitated the self-organisation of the cells, which improved the construct’s mechanical strength (compressive and tensile properties) and induced changes in glycosaminoglycan and type II collagen expression, resulting in improved tissue function. The compressive modulus of the construct reached the native cartilage range of 0.88 MPa in the 5th week of maturation. This paper reports the fabrication of anatomically sized and shaped cartilage constructs, achieved by combining novel iPSCs and bio-3D printers using a Kenzan needle array technology, which may facilitate chondral resurfacing of articular cartilage defects.

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Reemergence of undifferentiated cells from transplants of human induced pluripotent stem cells is a possible potential risk factor of tumorigenic differentiation

Cited by 4 publications

References 28 publications

Reversible transformation and de-differentiation of human cells derived from induced pluripotent stem cell teratomas

Reversible transformation and de-differentiation of human cells derived from induced pluripotent stem cell teratomas

Induced pluripotent stem cells for cartilage repair: current status and future perspectives

Bio-3D printing iPSC-derived human chondrocytes for articular cartilage regeneration

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