Cell Fusion Enhances Mesendodermal Differentiation of Human Induced Pluripotent Stem Cells

Qin, Jie; Sontag, Stephanie; Lin, Qiong; Mitzka, Saskia; Leisten, Isabelle; Schneider, Rebekka K.; Wang, Xiaoying; Jauch, Anna; Peitz, Michael; Brüstle, Oliver; Wagner, Wolfgang; Zhao, Robert Chunhua; Zenke, Martin

doi:10.1089/scd.2014.0120

Cited by 6 publications

(9 citation statements)

References 40 publications

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“…S1d and e, available at http://links.lww.com/PAIN/A749). HUES6 embryonal stem cells (ES cells) and iPS cells of healthy Caucasian non-IEM subjects (Ctrl 1 and Ctrl 2 33 ) were used as control.…”

Section: Resultsmentioning

confidence: 99%

“…Induced pluripotent stem cells derived from the 2 IEM patients are labelled IEM 1 (mother; EBiSC repository name UKAi0005-A) and IEM 2 (daughter; EBiSC repository name UKAi0006-A). HUES6 ES cells and iPS cells of healthy Caucasian non-IEM subjects (Ctrl 1, female, age 30 years, unpublished; Ctrl 2, female, age 3 years, previously referred to as iPS_2 33 ) were used as control. All experiments with human embryonic stem cells (HUES6) were performed in accordance with the German Stem Cell Act (RKI AZ.3.04.02/0085) and in the same manner as described below for iPS cells.…”

Section: Methodsmentioning

confidence: 99%

“…To assess 3 germ layer differentiation potential, iPS cells were subjected to spontaneous differentiation as described previously. 33 Briefly, embryoid bodies were formed by 30 minutes of collagenase IV (Thermo Fisher Scientific) treatment, transferred to ultralow attachment plates (Corning), and grown in suspension with KO-DMEM supplemented with 20% fetal calf serum (Lonza, Basel, Switzerland), 100 µM NEAA, 100 U/mL penicillin, 100 µg/mL streptomycin, 2 mM L-glutamine, and 0.1 mM β-mercaptoethanol. Samples were harvested for RNA isolation on day 0, 7, 14, 17, and 21 of differentiation and analyzed for germ layer–specific genes by RT-qPCR.…”

Section: Methodsmentioning

confidence: 99%

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The role of Nav1.7 in human nociceptors: insights from human induced pluripotent stem cell–derived sensory neurons of erythromelalgia patients

Meents

Bressan

Sontag

et al. 2019

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The role of Nav1.7 in human nociceptors: insights from human induced pluripotent stem cell–derived sensory neurons of erythromelalgia patients

Meents

Bressan

Sontag

et al. 2019

Pain

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“…As a standard line, we used H9 hESCs (H9) ( Thomson et al., 1998 ). Secondly, a tetraploid hybrid line generated by fusion of H9 hESCs and hematopoietic stem cells with a reported differentiation bias toward mesendoderm was used (H9Hyb) ( Qin et al., 2014 ). Thirdly, we generated hiPSCs (LU07) from skin fibroblasts using a polycistronic lentivirus with Dox-inducible transgenes OCT3/4 , SOX2 , KLF4 , and c-MYC ( Figure 1 A; Carey et al., 2009 ).…”

Section: Resultsmentioning

confidence: 99%

Differentiation-Defective Human Induced Pluripotent Stem Cells Reveal Strengths and Limitations of the Teratoma Assay and In Vitro Pluripotency Assays

et al. 2017

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SummaryThe ability to form teratomas in vivo containing multiple somatic cell types is regarded as functional evidence of pluripotency for human pluripotent stem cells (hPSCs). Since the Teratoma assay is animal dependent, laborious, and only qualitative, the PluriTest and the hPSC ScoreCard assay have been developed as in vitro alternatives. Here we compared normal hPSCs, induced hPSCs (hiPSCs) with reactivated reprogramming transgenes, and human embryonal carcinoma cells (hECs) in these assays. While normal hPSCs gave rise to typical teratomas, the xenografts of the hECs and the hiPSCs with reactivated reprogramming transgenes were largely undifferentiated and malignant. The hPSC ScoreCard assay confirmed the line-specific differentiation propensities in vitro. However, when undifferentiated cells were analyzed by the PluriTest, only hECs were identified as abnormal whereas all other cell lines were indistinguishable and resembled normal hPSCs. Our results indicate that pluripotency assays are best selected on the basis of intended downstream applications.

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“…HSCs [30]. Nevertheless, concerns remain that iPSCs possess inherent defects in multiple functional lymphocyte lineages, which may hinder their advantages in the clinical treatment of hematological disorders.…”

Section: Introductionmentioning

confidence: 99%

Induced Pluripotent Stem Cells Can Effectively Differentiate into Multiple Functional Lymphocyte Lineages In Vivo with Negligible Bias

Lan

Wang

et al. 2016

Stem Cells and Development

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Lymphohematopoietic stem cells (L-HSCs) generated from self-somatic cell-derived induced pluripotent stem cells (iPSCs) are a potential source of cells for the treatment of hematological disorders. However, the generation of truly functional L-HSCs from iPSCs has yet to be achieved. Thus, whether iPSCs have the inherent potential to generate a normal differentiated phenotype and functional population of multiple lineages of terminally differentiated lymphocytes needs to be assessed. Here, we used tetraploid embryo complementation to provide a normal environment for the differentiation of hematopoietic cells from iPSCs and embryonic stem cells (ESCs). We then evaluated the characteristics, populations, and functions of lymphocytes derived from iPSCs, ESCs, and naïve isogenic C57BL/6 mice. The results showed that iPSC-derived lymphocytes (iPSLs) expressed normal levels of major histocompatibility complex-I (MHC-I) and exhibited a fully pluripotent capacity to differentiate into CD4(+) T, CD8(+) T, regulatory T, B, and natural killer cells. Following in vitro stimulation with either concanavalin A or an alloantigen, iPSLs exhibited the same capacities for proliferation and cytokine secretion as ESC-derived or isogenic lymphocytes. Furthermore, iPSC-derived bone marrow cells could differentiate into multiple lymphocyte lineages that reconstituted the lymphocyte population in syngeneic lethally irradiated recipient animals. Our results demonstrated that iPSCs have the inherent potential to differentiate into multiple lineages of functional lymphocytes without bias, and further support the practical application of iPSC-based treatments to hematological disorders.

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Cell Fusion Enhances Mesendodermal Differentiation of Human Induced Pluripotent Stem Cells

Cited by 6 publications

References 40 publications

The role of Nav1.7 in human nociceptors: insights from human induced pluripotent stem cell–derived sensory neurons of erythromelalgia patients

The role of Nav1.7 in human nociceptors: insights from human induced pluripotent stem cell–derived sensory neurons of erythromelalgia patients

Differentiation-Defective Human Induced Pluripotent Stem Cells Reveal Strengths and Limitations of the Teratoma Assay and In Vitro Pluripotency Assays

Induced Pluripotent Stem Cells Can Effectively Differentiate into Multiple Functional Lymphocyte Lineages In Vivo with Negligible Bias

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