Current methods of adipogenic differentiation of mesenchymal stem cells

Abstract

There has been a recent increase in our understanding in the isolation, culture, and differentiation of mesenchymal stem cells (MSCs). Concomitantly, the availability of MSCs has increased, with cells now commercially available, including human MSCs from adipose tissue and bone marrow. Despite an increased understanding of MSC biology and an increase in their availability, standardization of techniques for adipogenic differentiation of MSCs is lacking. The following review will explore the variability in adipogenic differentiation in vitro, specifically in 3T3-L1 and primary MSCs derived from both adipose tissue and bone marrow. A review of alternative methods of adipogenic induction is also presented, including the use of specific peroxisome proliferator-activated receptor-gamma agonists as well as bone morphogenetic proteins. Finally, we define a standard, commonly used adipogenic differentiation medium in the hopes that this will be adopted for the future standardization of laboratory techniques--however, we also highlight the essentially arbitrary nature of this decision. With the current, rapid pace of electronic publications, it becomes imperative for standardization of such basic techniques so that interlaboratory results may be easily compared and interpreted.

FIG. 1.

Variation in adipogenic differentiation of 3T3-L1 cells. A literature review for articles within 2010 was performed for any publication examining the adipogenic differentiation of 3T3-L1 preadipocytes. (A) Breakdown of 45 publications by country of origin. (B) Breakdown of publications by area of scientific interest. (C) Breakdown of 3 major components of induction medium used for each individual publication. Although nearly all publications used insulin, dexamethasone, and IBMX, the concentrations varied widely. See Table 2 for a complete listing of induction components.

FIG. 2.

Variation in adipogenic differentiation of BMSCs. Again, a literature review for articles within 2010 was performed for any publication examining the adipogenic differentiation of BMSCs—broken down by either mouse (left) or human (right) origin. (A) Country of origin for each article. (B) Area of scientific interest. (C) Breakdown of 3 major components of induction medium used for each individual publication. Although most publications used insulin, dexamethasone, and IBMX, the concentrations varied widely. See Tables 3 and 4 for a complete listing of induction components. BMSCs, bone marrow mesenchymal stem cells.

FIG. 3.

Differences in adipogenic differentiation of BMSCs and ASCs based on species. Median values for each component of BMSC/ASC adipogenic induction medium was calculated and compared between mouse and human cells. (A) Mean concentration of insulin. (B) Mean concentration of dexamethasone. (C) Mean concentration of IBMX. ASCs, adipose-derived mesenchymal stem cells.

FIG. 4.

Variation in adipogenic differentiation of ASCs. Again, a literature review for articles within 2010 was performed for any publication examining the adipogenic differentiation of ASCs—broken down by either mouse (left) or human (right) origin. (A) Country of origin. (B) Area of scientific interest. (C) Breakdown of 4 major components of induction medium used for each individual publication. Although most publications used indomethacin, insulin, dexamethasone, and IBMX, the concentrations varied widely.