scholarly journals The Construction and Identification of Induced Pluripotent Stem Cells Derived from Acute Myelogenous Leukemia Cells

2017 ◽  
Vol 41 (4) ◽  
pp. 1661-1674 ◽  
Author(s):  
Liang-Fang Zhu ◽  
Qiu-Ru Chen ◽  
Shao-Zhen Chen ◽  
Ling-Yan Wang ◽  
Xiao-Feng Luo ◽  
...  
Keyword(s):  
Embryoid Body ◽  
Karyotype Analysis ◽  
Myelogenous Leukemia ◽  
Ipsc Line ◽  
Body Formation ◽  
Qrt Pcr ◽  
Acute Myelogenous ◽  
Induced Pluripotent ◽  
Teratoma Formation ◽  
Embryoid Body Formation

Objective: The present study aimed to establish an induced pluripotent stem cell (iPSC) line from acute myelogenous leukemia (AML) cells in vitro and identify their biological characteristics. Methods: Cells from the AML-infiltrated skin from an M6 patient were infected with a lentivirus carrying OCT4, SOX2, KLF4 and C-MYC to induce iPSCs. The characteristics of the iPSCs were confirmed by alkaline phosphatase (ALP) staining. The proliferation ability of iPSCs was detected with a CCK-8 assay. The expression of pluripotency markers was measured by immunostaining, and the expression of stem cell-related genes was detected by qRT-PCR; distortion during the induction process was detected by karyotype analysis; the differentiation potential of iPSCs was determined by embryoid body-formation and teratoma-formation assays. ALP staining confirmed that these cells exhibited positive staining and had the characteristics of iPSCs. Results: The CCK-8 assay showed that the iPSCs had the ability to proliferate. Immunostaining demonstrated that iPSC clones showed positive expression of NANOG, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81. qRT-PCR results revealed that the mRNA expression of Nanog, Lin28, Cripto, FOX3, DNMT3b, DPPA2, and DPPA4 significantly increased in iPSCs. Karyotype analysis found no chromosome aberration in the iPSCs. The results of the embryoid body-formation and teratoma-formation assays indicated that the iPSCs had the potential to differentiate into all three germ layers. Conclusion: Our study provided evidence that an iPSC line derived from AML cells was successfully established.

Effective embryoid body formation from induced pluripotent stem cells for regeneration of respiratory epithelium

2013 ◽  
Vol 124 (1) ◽  
pp. E8-E14 ◽  
Author(s):  
Koshi Otsuki ◽  
Mitsuyoshi Imaizumi ◽  
Yukio Nomoto ◽  
Mika Nomoto ◽  
Ikuo Wada ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Embryoid Body ◽  
Respiratory Epithelium ◽  
Body Formation ◽  
Induced Pluripotent ◽  
Embryoid Body Formation

scholarly journals Differentiation of Induced Pluripotent Stem Cells into Male Germ CellsIn Vitrothrough Embryoid Body Formation and Retinoic Acid or Testosterone Induction

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Peng Li ◽  
Hongliang Hu ◽  
Shi Yang ◽  
Ruhui Tian ◽  
Zhenzhen Zhang ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Embryoid Body ◽  
Ips Cells ◽  
Male Germ Cells ◽  
Body Formation ◽  
Induced Pluripotent ◽  
Embryoid Body Formation

Generation of germ cells from pluripotent stem cellsin vitrocould have great application for treating infertility and provides an excellent model for uncovering molecular mechanisms controlling gametogenesis. In this study, we explored the differentiation potential of mouse induced pluripotent stem (iPS) cells towards male germ cells. Embryoid body formation and retinoic acid/testosterone induction were applied to promote differentiation of mouse iPS cells into male germ cellsin vitro. Quantitative RT-PCR and immunoflourescence were performed to characterize the iPS cell differentiation process, and notably there were different temporal expression profiles of male germ cell-associated genes. The expression of proteins, including MVH, CDH1, and SCP3, was remarkably increased. mRNA expression ofStra8,Odf2,Act, andPrm1was upregulated in iPS cells by retinoic acid or testosterone induction, whereasOct-4transcription was reduced in these cells compared to the controls. Hormones were also measured in the EB medium. DNA content analysis by flow cytometry revealed that iPS cells could differentiate into haploid cells through retinoic acid or testosterone treatment. Collectively, our results suggest that mouse iPS cells possess the potency to differentiate into male germ cellsin vitrothrough embryoid body formation and retinoic acid or testosterone induction.

scholarly journals Single embryoid body formation in a multi-well plate

2007 ◽  
Vol 10 (2) ◽  
pp. 0-0 ◽  
Author(s):  
Uthayashanker R. Ezekiel ◽  
Mariappan Muthuchamy ◽  
Jan S. Ryerse ◽  
Rita M. Heuertz
Keyword(s):  
Embryoid Body ◽  
Body Formation ◽  
Embryoid Body Formation

Methods for Embryoid Body Formation: The Microwell Approach

2010 ◽  
pp. 151-162 ◽  
Author(s):  
Dawn P. Spelke ◽  
Daniel Ortmann ◽  
Ali Khademhosseini ◽  
Lino Ferreira ◽  
Jeffrey M. Karp
Keyword(s):  
Embryoid Body ◽  
Body Formation ◽  
Embryoid Body Formation
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