Tumorigenic risk of human induced pluripotent stem cell explants cultured on mouse SNL76/7 feeder cells

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

doi:10.1016/j.bbrc.2014.10.009

Cited by 8 publications

(12 citation statements)

References 14 publications

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“…We found that any clones did not express the neomycin resistant gene ( Neo r ) (Fig. 1 e), confirming that the cells were derived from hiPSCs but not mouse feeder cells, as aforementioned [ 9 ]. The fibroblast secretary protein-1 ( FSP - 1 ) gene was expressed in all the clones, suggesting that they were not epithelial cells.…”

Section: Resultssupporting

confidence: 81%

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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: Resultssupporting

confidence: 81%

“…The reason was not explained well [ 5 ]. Our finding on formation of various malignant tumors due to contaminated mouse feeder cells in the explant might explain some of the reasons [ 9 ]. However, unknown mechanisms seem to exist in emergence of transformed cells from iPSC derivatives.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2015

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“…Promising cells are also iPSCs that have shown the ability to differentiate into motoneurons in vitro [57,58,59] and to form functional connections with reinnervated muscle in vivo [26]. Embryonic fetal cells are safer compared to IPSCs because there is no danger of virus activation or cancer development due to induced pluripotency [60,61,62,63,64,65]. In addition, differentiation potential of iPSCs into specific motoneurons is not well characterized and it is very difficult to obtain fully mature motoneurons from iPSCs [66].…”

Section: Discussionmentioning

confidence: 99%

Transplantation of Embryonic Spinal Cord Derived Cells Helps to Prevent Muscle Atrophy after Peripheral Nerve Injury

Ruven

et al. 2017

IJMS

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Injuries to peripheral nerves are frequent in serious traumas and spinal cord injuries. In addition to surgical approaches, other interventions, such as cell transplantation, should be considered to keep the muscles in good condition until the axons regenerate. In this study, E14.5 rat embryonic spinal cord fetal cells and cultured neural progenitor cells from different spinal cord segments were injected into transected musculocutaneous nerve of 200–300 g female Sprague Dawley (SD) rats, and atrophy in biceps brachii was assessed. Both kinds of cells were able to survive, extend their axons towards the muscle and form neuromuscular junctions that were functional in electromyographic studies. As a result, muscle endplates were preserved and atrophy was reduced. Furthermore, we observed that the fetal cells had a better effect in reducing the muscle atrophy compared to the pure neural progenitor cells, whereas lumbar cells were more beneficial compared to thoracic and cervical cells. In addition, fetal lumbar cells were used to supplement six weeks delayed surgical repair after the nerve transection. Cell transplantation helped to preserve the muscle endplates, which in turn lead to earlier functional recovery seen in behavioral test and electromyography. In conclusion, we were able to show that embryonic spinal cord derived cells, especially the lumbar fetal cells, are beneficial in the treatment of peripheral nerve injuries due to their ability to prevent the muscle atrophy.

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“…Previous studies, including our own, have reported that human iPSC-derived peripheral neurons [ 5 , 6 ], melanocytes [ 7 ], and MSCs [ 8 ] are generated via the neural crest cell lineage. iPSCs possess numerous advantages such as infinite self-renewability and differentiation potential, however, they also harbor the potential risks of tumorigenicity and differentiation uncertainty [ 9 , 10 ]. This concern led to the research for alternative cell sources and cell induction approaches.…”

Section: Introductionmentioning

confidence: 99%

Exposure to small molecule cocktails allows induction of neural crest lineage cells from human adipose-derived mesenchymal stem cells

et al. 2020

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Neural crest cells (NCCs) are a promising source for cell therapy and regenerative medicine owing to their multipotency, self-renewability, and capability to secrete various trophic factors. However, isolating NCCs from adult organs is challenging, because NCCs are broadly distributed throughout the body. Hence, we attempted to directly induce NCCs from human adipose-derived mesenchymal stem cells (ADSCs), which can be isolated easily, using small molecule cocktails. We established a controlled induction protocol with two-step application of small molecule cocktails for 6 days. The induction efficiency was evaluated based on mRNA and protein expression of neural crest markers, such as nerve growth factor receptor (NGFR) and sex-determining region Y-box 10 (SOX10). We also found that various trophic factors were significantly upregulated following treatment with the small molecule cocktails. Therefore, we performed global profiling of cell surface makers and identified distinctly upregulated markers, including the neural crest-specific cell surface markers CD271 and CD57. These results indicate that our chemical treatment can direct human ADSCs to developing into the neural crest lineage. This offers a promising experimental platform to study human NCCs for applications in cell therapy and regenerative medicine.

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Tumorigenic risk of human induced pluripotent stem cell explants cultured on mouse SNL76/7 feeder cells

Cited by 8 publications

References 14 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

Transplantation of Embryonic Spinal Cord Derived Cells Helps to Prevent Muscle Atrophy after Peripheral Nerve Injury

Exposure to small molecule cocktails allows induction of neural crest lineage cells from human adipose-derived mesenchymal stem cells

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