Generation of novel adipocyte monolayer cultures from embryonic stem cells

Stem Cells and Development
Theresa L ChenFredric B Kraemer

Abstract

Here we show a simplified and improved method to produce large quantities of evenly distributed monolayer cultures that display major characteristics of adipocytes. These cultures are applicable for quantitative analysis for biochemical and molecular events in adipogenesis during development and may provide a useful system for high-throughput drug screening assays of antiobesity drugs. In our method, we treated embryoid bodies (EBs) with all-trans retinoic acid (ATRA) for 3 days, 1 day after they attached to the gelatin-coated culture plates without further transfer. The cells were maintained in insulin and trioiodothyronine (T(3))-containing medium until day 12, when they were dispersed by enzymatic digestion and replated onto multiple culture plates. Two days later, adipocyte induction factors were added for 6 days and examined 6 days later. The amount of lipid droplet-laden adipocytes in the culture reached approximately 80%, with a nearly five-fold increase in GPDH activity. The cells expressed high levels of adipose-specific proteins (adipocyte markers), including PPARgamma2, ALBP, LPL, HSL, perilipin, and DGAT1. The adipocytes are functionally active, as evidenced by their response to lipolytic agents, such as forskolin, ...Continue Reading

References

Jul 1, 1992·Histochemistry·J L Ramírez-ZacaríasWalid Kuri-Harcuch
Dec 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·G R Martin
Apr 1, 1995·Developmental Biology·G BainD I Gottlieb
Aug 1, 1996·Biology of Reproduction·J A ThomsonJ P Hearn
Nov 6, 1998·Science·J A ThomsonJ M Jones
May 13, 1999·Proceedings of the National Academy of Sciences of the United States of America·W J ShenFredric B Kraemer
May 20, 1999·The Anatomical Record·K S O'Shea
Aug 13, 1999·Molecular Cell·L J Syu, A R Saltiel
Dec 11, 1999·Cells, Tissues, Organs·J RohwedelAnna M Wobus
Oct 12, 2000·Proceedings of the National Academy of Sciences of the United States of America·M SchuldinerN Benvenisty
Mar 13, 2001·International Journal of Obesity and Related Metabolic Disorders : Journal of the International Association for the Study of Obesity·M WabitschH Hauner
Jun 8, 2001·Biochemical and Biophysical Research Communications·B W PhillipsChristian Dani
Oct 5, 2002·Journal of Lipid Research·Fredric B Kraemer, Wen-Jun Shen
Jan 14, 2003·Nature Biotechnology·Qi-Long YingAustin G Smith
Feb 1, 2003·Differentiation; Research in Biological Diversity·Nicole I zur NiedenHans J Ahr
Aug 28, 2003·The Journal of Biological Chemistry·Wu YinMorris J Birnbaum
Nov 1, 2003·Biochemical and Biophysical Research Communications·Mikael RydénKatarina Le Blanc
Nov 19, 2003·Biochemical and Biophysical Research Communications·Enrique Juárez-AguilarWalid Kuri-Harcuch
Mar 25, 2005·Physiological Reviews·Anna M Wobus, Kenneth R Boheler
Jan 26, 2006·Stem Cells and Development·Chen XiongYuqing E Chen
Mar 31, 2006·Cloning and Stem Cells·Akihiro YamashitaRyuzo Torii

Citations

Sep 4, 2008·Stem Cells and Development·Nicholas R F Hannan, Ernst J Wolvetang
May 14, 2011·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Wen-Jun ShenFredric B Kraemer
May 27, 2014·Stem Cell Research·Ixchelt Cuaranta-MonroyLaszlo Nagy

Related Concepts

Antineoplastic Agents
Cell Differentiation Process
Bucladesine, Barium (1: 1) Salt
Dose-Response Relationship, Drug
Colforsin
Vesanoid
Viral Markers
Adipocytes
Cell Culture Techniques
Adipogenesis

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Synthetic Genetic Array Analysis

Synthetic genetic arrays allow the systematic examination of genetic interactions. Here is the latest research focusing on synthetic genetic arrays and their analyses.

Congenital Hyperinsulinism

Congenital hyperinsulinism is caused by genetic mutations resulting in excess insulin secretion from beta cells of the pancreas. Here is the latest research.

Neural Activity: Imaging

Imaging of neural activity in vivo has developed rapidly recently with the advancement of fluorescence microscopy, including new applications using miniaturized microscopes (miniscopes). This feed follows the progress in this growing field.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Epigenetic Memory

Epigenetic memory refers to the heritable genetic changes that are not explained by the DNA sequence. Find the latest research on epigenetic memory here.

Cell Atlas of the Human Eye

Constructing a cell atlas of the human eye will require transcriptomic and histologic analysis over the lifespan. This understanding will aid in the study of development and disease. Find the latest research pertaining to the Cell Atlas of the Human Eye here.

Femoral Neoplasms

Femoral Neoplasms are bone tumors that arise in the femur. Discover the latest research on femoral neoplasms here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.