In this unit standard culture conditions for mouse embryonic stem cells (mESCs) on primary murine embryonic fibroblast (PMEF or MEF) monolayers, culture conditions without MEF for feeder-independent mESCs, and culture conditions in chemically defined media for both feeder-independent mESCs and feeder-dependent mESCs are described. For expansion of an mESC line, it is crucial that cells maintain their undifferentiated state and their self-renewal capacity, and that they remain karyotypically normal, all of which are necessary for successful chimerization of the germ line upon blastocyst injection. Derivation and culture conditions for the original mESCs have been described (notably Robertson, 1987; Smith, 1991; Nagy et al., 2003), however, as there are more and more mESC lines available, it becomes evident that culture conditions are cell-line specific to some extent, and there is a constant demand for culturing details for mESC lines derived from different mouse strains.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1002/9780470151808.sc01c04s5 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Preparing the 2025 revision of the International Code of Nomenclature of Prokaryotes.
Int J Syst Evol Microbiol
January 2025
Department of Infectious Disease, Institute for Biomedicine, Sahlgrenska Academy, University of Gothenburg, P.O. Box 7193, SE-402 34 Gteborg, Sweden.
The editorial Board of the (ICNP) - the Prokaryotic Code - has compiled already ratified proposed emendations of the ICNP, together with additional editorial changes and clarifications. These were implemented in a draft 2025 revision of the . To comply with Articles 13(b)(4) and 4(d) of the statutes of the International Committee on Systematics of Prokaryotes (ICSP), a public discussion of the document will start on 1 January (or later if required) 2025, to last for 6 months.
View Article and Find Full Text PDFRacial and Ethnic Disparities in Pediatric Counseling on Nutrition, Lifestyle, and Weight: A Secondary Analysis of the BP-CATCH Randomized Clinical Trial.
JAMA Netw Open
January 2025
Department of Pediatrics, The Children's Hospital at Montefiore, Albert Einstein College of Medicine, Bronx, New York.
Importance: Pediatric obesity and hypertension are highly correlated. To mitigate both conditions, provision of counseling on nutrition, lifestyle, and weight to children with high blood pressure (BP) measurements is recommended.
Objective: To examine racial and ethnic disparities in receipt of nutrition, lifestyle, and weight counseling among patients with high BP at pediatric primary care visits stratified by patients' weight status.
Quantitative Analysis of Mitochondria-Associated Endoplasmic Reticulum Membrane (MAM) Stabilization in a Neural Model of Alzheimer's Disease (AD).
J Vis Exp
January 2025
Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Henry and Allison McCance Center for Brain Health, Department of Neurology, Massachusetts General Hospital, Harvard Medical School;
A method to quantitate the stabilization of Mitochondria-Associated endoplasmic reticulum Membranes (MAMs) in a 3-dimensional (3D) neural model of Alzheimer's disease (AD) is presented here. To begin, fresh human neuro progenitor ReN cells expressing β-amyloid precursor protein (APP) containing familial Alzheimer's disease (FAD) or naïve ReN cells are grown in thin (1:100) Matrigel-coated tissue culture plates. After the cells reach confluency, these are electroporated with expression plasmids encoding red fluorescence protein (RFP)-conjugated mitochondria-binding sequence of AKAP1(34-63) (Mito-RFP) that detects mitochondria or constitutive MAM stabilizers MAM 1X or MAM 9X that stabilize tight (6 nm ± 1 nm gap width) or loose (24 nm ± 3 nm gap width) MAMs, respectively.
View Article and Find Full Text PDFFoxO1 promotes high glucose-induced inflammation and cataract formation via JAK1/STAT1.
Graefes Arch Clin Exp Ophthalmol
January 2025
National Clinical Research Center for Ocular Diseases, Eye Hospital, Wenzhou Medical University, 270 Xueyuan West Road, Wenzhou, 325027, Zhejiang, China.
Purpose: To investigate whether in diabetic cataract (DC), FoxO1 regulates high glucose (HG)-induced activation of NLRC4/IL-6 inflammatory mediators in human lens epithelial cells (SRA01/04) via the JAK1/STAT1 pathway, leading to cataract formation.
Methods: Expression levels of FoxO1, inflammatory factor IL-6 and inflammatory vesicle NLRC4 were examined in SRA01/04 under high glucose (HG) stress at 25-150 mM. Rat lenses were also cultured using HG medium with or without the addition of the FoxO1 inhibitor AS1842856 and the JAK1 agonist RO8191.
Racial Differences in Discrepancies Between Subjective Ratings and Objective Assessments of Hearing Impairment.
Gerontologist
January 2025
School of Aging Studies, University of South Florida, Tampa, Florida, USA.
Background And Objectives: To better understand racial/ethnic disparities in hearing aid use, we examined racial differences in discrepancies between subjective hearing ratings and objective hearing tests as a potential source of this disparity.
Research Design And Methods: A cross-sectional assessment was conducted using the data from the Health and Retirement Study (HRS). Our analytic sample included 2,568 participants aged 50 and older: 1,814 non-Hispanic White Americans and 754 non-Hispanic Black Americans.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!