Indentifying early morphological transitions after osteogenic and adipogenic induction of murine embryonic stem cells

Tuesday, October 09, 2012 — Poster Session I
1:00 p.m. – 3:00 p.m	Natcher Conference Center, Building 45	NIA	AGING-3

Authors

• J.D. Delaney
• C.E. Coletta
• D.M. Eckley
• N. Orlov
• I.G. Goldberg

Abstract

In aging the bone remodeling process becomes perturbed favoring fat generation over new bone deposition. To gain a better understanding of the regulatory switches involved, mESC were treated with osteogenic (OS) or adipogenic (AD) media, imaged for 7 days (phase contrast microscopy) and analyzed with a pattern recognition approach to image processing (WND-CHARM). Calculated distances between the OS induced cells and controls demonstrated an abrupt increase in distance between day 2 (5.77u) and day 3 (12.45u). Cells treated with AD also demonstrate an abrupt change occurring between day 3 (6.75u) and day 4 (12.65u). These morphologic profiles coincide with rtPCR data demonstrating osteogenic (BSP: 9 to 14.6 and Osterix: 0.21 to 8.0) and adipogenic (AP2 8.51 to 38.20 and PPAR-g 4.02 to 35.63) commitment respectively. Negative controls demonstrate very little adipogenic activation in osteogenic cultures (AP2: 0.1 to 2.70 and PPAR-g 2.12 to 0.00) and a decrease in osteogenic activation in adipogenic cultures (BSP: 13.04 to 0.42 and Osterix: 2.07 to 0.03). Here we were able to demonstrate that the transition state predicted by WNDCHARM coincides with an actual molecular switch. In the future these switch points will be more finely refined with micro-array analysis.

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