Pluripotent Differentiation in vitro of Murine ES-D3 Embryonic Stem Cells

Toumadje, Arazdordi; Kusumoto, Ken-Ichi; Parton, Angela; Mericko, Patricia; Dowell, Lori; Ma, Guozhong; Chen, Luping; Barnes, David; Sato, J. Denry

doi:10.1290/0309071

Cited by 16 publications

(17 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1B), indicating multiple interferences on the cell cycle at this condition. We also tested the effects of Olo II on D3 cells, a well-characterized and commonly used mESC line by other investigators [27]. It is noted that the reduction of the G1 phase cells in D3 cells is less dramatic and the reduction in S phase populations is more obvious than that observed in DBA252 cells at the concentrations of 5 μM and 10 μM, however, the accumulation of G2 cells is similar between the two cell lines.…”

Section: Resultsmentioning

confidence: 96%

Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

Wang

Guo

2012

Experimental Cell Research

View full text Add to dashboard Cite

Embryonic stem cells (ESCs) have unlimited capacity for self-renewal and can differentiate into various cell types when induced. They also have an unusual cell cycle control mechanism driven by constitutively active cyclin dependent kinases (Cdks). In mouse ESCs (mESCs). It is proposed that the rapid cell proliferation could be a necessary part of mechanisms that maintain mESC self-renewal and pluripotency, but this hypothesis is not in line with the finding in human ESCs (hESCs) that the length of the cell cycle is similar to differentiated cells. Therefore, whether rapid cell proliferation is essential for the maintenance of mESC state remains unclear. We provide insight into this uncertainty through chemical intervention of mESC cell cycle. We report here that inhibition of Cdks with olomoucine II can dramatically slow down cell proliferation of mESCs with concurrent down-regulation of cyclin A, B and E, and the activation of the Rb pathway. However, mESCs display can recover upon the removal of olomoucine II and are able to resume normal cell proliferation without losing self-renewal and pluripotency, as demonstrated by the expression of ESC markers, colony formation, embryoid body formation, and induced differentiation. We provide a mechanistic explanation for these observations by demonstrating that Oct4 and Nanog, two major transcription factors that play critical roles in the maintenance of ESC properties, are up-regulated via de novo protein synthesis when the cells are exposed to olomoucine II. Together, our data suggest that short-term inhibition of cell proliferation does not compromise the basic properties of mESCs.

show abstract

Section: Resultsmentioning

confidence: 96%

Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

Wang

Guo

2012

Experimental Cell Research

View full text Add to dashboard Cite

show abstract

“…The mESC were maintained in an undifferentiated state on a feeder layer of irradiated mouse fibroblasts with maintenance medium containing Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, CA, USA) supplemented with 15% FBS (Hyclone, Logan, UT), 0.1 mM 2‐mercaptoethanol (Sigma, St. Louis, MO, USA), 0.1 mN non‐essential amino acids (Invitrogen), 1% penicillin/streptomycin (Invitrogen), 2 mM L‐glutamine (Invitrogen), and 1000 U/mL mouse leukemia inhibitory factor (LIF, ESGRO; Chemicon, Temecula, CA, USA) at 37°C as described previously (12). Three stable lines of mESC were employed: 1) the TL‐1 mESC line derived from 129sv/J mice (courtesy of Dr. Thomas Quertermous, Stanford University, Stanford, CA), 2) luciferase‐transfected mESC (mESC‐luc+, courtesy of Dr. Joseph C. Wu, Stanford University, Stanford, CA), and 3) Nkx2.5 (cardiomyocyte differentiation specific promoter)‐green fluorescent protein (GFP) transfected mESC (mESC‐Nkx2.5‐GFP + , courtesy of Dr. Takayuki Morisaki, National Cardiovascular Center Research Institute, Osaka, Japan) were used.…”

Section: Methodsmentioning

confidence: 99%

In vitro comparison of the biological effects of three transfection methods for magnetically labeling mouse embryonic stem cells with ferumoxides

Suzuki¹,

Zhang²,

Kundu³

et al. 2007

Magnetic Resonance in Med

View full text Add to dashboard Cite

In vivo MRI of stem cells (SCs) is an emerging application to evaluate the role of cell therapy in restoring the injured myocardium. The high spatial and temporal resolution combined with iron-oxide-based intracellular labeling techniques will provide a sensitive, noninvasive, dual imaging modality for both cells and myocardium. In order to facilitate this novel imaging approach, much effort has been directed towards developing efficient transfection methods. While techniques utilizing poly-L-lysine (PLL), protamine sulfate (PS), and electroporation (ELP) have been proposed, the fundamental biological effects of these methods on mouse embryonic SCs (mESC) have not been investigated systematically. In this study a longitudinal in vitro evaluation of cellular viability, apoptosis, proliferation, and cardiac differentiation of magnetically labeled mESC was conducted. No significant difference was seen in these biological parameters among the three transfection methods. However, cardiac differentiation was most attenuated by ELP, and iron Growing evidence from preclinical and clinical studies suggests that cell therapy can improve cardiac function (1-4). Although this therapeutic option is a promising approach to repair the injured myocardium, the exact mechanism of functional restoration of the injured myocardium is unknown. While differentiation of embryonic stem cells (ESC) into functional cardiomyocytes has been attributed as one of the possible underlying mechanisms in restoring the injured myocardium, a more fundamental issue regarding cellular viability following delivery into the myocardium also needs to be addressed (5,6). At present, most cell therapy protocols require histological analysis to determine viable engraftment of the transplanted cells. The development of sensitive, noninvasive technologies to monitor this fundamental engraftment parameter will aid clinical implementation of cell therapy. Imaging modalities such as single-photon emission computed tomography (SPECT), positron emission tomography (PET), magnetic resonance imaging (MRI), and optical bioluminescence imaging (BLI) have been reported to be capable of evaluating cellular engraftment parameters (4,7,8).In vivo MRI of SCs is an emerging application to monitor SC engraftment. The high spatial and temporal resolution combined with robust iron-oxide-based intracellular labeling enables dual assessment of myocardial function and cellular location. Much effort has been directed towards developing efficient transfection methods to facilitate intracellular magnetic labeling with superparamagnetic iron oxide (SPIO). Although techniques utilizing poly-L-lysine (PLL), protamine sulfate (PS), and electroporation (ELP) have been proposed, the effects of these three transfection methods on engraftment parameters of ESC have not been investigated systematically (9 -11). In this study a comparison of the engraftment parameters of magnetically labeled mouse embryonic SCs (mESC) using the three transfection methods was conducted. Fundamental biological ...

show abstract

“…LIF has been applied not only for feeder cell-free culture but also for coculture with supporting cells, but its concentration varies according to culture systems (19)(20)(21)(22). Among the concentrations of LIF that have been added to culture media, 1,000 U/mL is optimal, regardless of the presence or absence of feeder cells (23)(24)(25)(26).…”

Section: Discussionmentioning

confidence: 99%

Requirement of leukemia inhibitory factor for establishing and maintaining embryonic stem cells in mice

Lee

et al. 2009

Fertility and Sterility

View full text Add to dashboard Cite

Pluripotent Differentiation in vitro of Murine ES-D3 Embryonic Stem Cells

Cited by 16 publications

References 0 publications

Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

In vitro comparison of the biological effects of three transfection methods for magnetically labeling mouse embryonic stem cells with ferumoxides

Requirement of leukemia inhibitory factor for establishing and maintaining embryonic stem cells in mice

Contact Info

Product

Resources

About