Presentation Title

Cited2 is a Negative Regulator of Human Adipogenesis

Faculty Mentor

Yuanxiang Ansel Zhao

Start Date

18-11-2017 2:15 PM

End Date

18-11-2017 3:15 PM

Location

BSC-Ursa Minor 83

Session

Poster 3

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

CITED2 Is a Negative Regulator of Human Adipogenesis

Human adipogenesis is the process through which uncommitted human mesenchymal stem cells (hMSCs) differentiate into adipocytes (fat cells). HMSCs normally reside in the bone marrow and adipose tissue. They can be easily isolated, expanded in vitro and upon exposure to a cocktail (AIM) of isobutylmethylxanthine (IBMX), Dexamethasone (DEX) and Insulin, can differentiate into mature adipocytes (fat cells), making it an excellent in vitro cellular model for studying human adipogenesis. Based on a high throughput screen using siRNAs targeting 5000 genes in the human genome, a list of genes whose expression knock-down by its siRNAs led to enhanced adipogenic differentiation of hMSCs were uncovered. This study has focused on one of the uncovered genes, CITED2, whose role in human adipogenesis has never been studied. Here we report that expression of CITED2 is upregulated by about 2-4 fold during normal adipogenic commitment stage, between day 3 to day 6 post adipogenic induction by AIM. Expression knockdown by siCITED2 led to about 80% reduction of its normal expression level, but resulted in near 2-fold increase in total fat accumulation based on OilRedO staining and extraction. To find out whether the increased fat accumulation was due to increased total fat cell numbers or increased fat accumulation in individual fat cells, total cell numbers and mature fat cells were counted, and the resulting percentage of fat cells was found to be significantly higher in siCITED2 treated group (25%) vs. siCONTROL treated group (7%). In addition, the total number of cells in both treatment groups were insignificantly different, indicating that expression knockdown of CITED2 during adipogenesis significantly promoted adipogenic differentiation efficiency of hMSCs, hence increasing the total number of mature fat cells. Furthermore, real time RT-PCR analysis demonstrated that the expression of CEBPα and PPARγ, two master regulators of adipogensis, were both dramatically increased in siCITED2 cells compare to siControl cells, suggesting that CITED2 normally acts as negative regulator of human adipogenesis by suppressing the expression of CEBPα and PPARγ.

This document is currently not available here.

Share

COinS
 
Nov 18th, 2:15 PM Nov 18th, 3:15 PM

Cited2 is a Negative Regulator of Human Adipogenesis

BSC-Ursa Minor 83

CITED2 Is a Negative Regulator of Human Adipogenesis

Human adipogenesis is the process through which uncommitted human mesenchymal stem cells (hMSCs) differentiate into adipocytes (fat cells). HMSCs normally reside in the bone marrow and adipose tissue. They can be easily isolated, expanded in vitro and upon exposure to a cocktail (AIM) of isobutylmethylxanthine (IBMX), Dexamethasone (DEX) and Insulin, can differentiate into mature adipocytes (fat cells), making it an excellent in vitro cellular model for studying human adipogenesis. Based on a high throughput screen using siRNAs targeting 5000 genes in the human genome, a list of genes whose expression knock-down by its siRNAs led to enhanced adipogenic differentiation of hMSCs were uncovered. This study has focused on one of the uncovered genes, CITED2, whose role in human adipogenesis has never been studied. Here we report that expression of CITED2 is upregulated by about 2-4 fold during normal adipogenic commitment stage, between day 3 to day 6 post adipogenic induction by AIM. Expression knockdown by siCITED2 led to about 80% reduction of its normal expression level, but resulted in near 2-fold increase in total fat accumulation based on OilRedO staining and extraction. To find out whether the increased fat accumulation was due to increased total fat cell numbers or increased fat accumulation in individual fat cells, total cell numbers and mature fat cells were counted, and the resulting percentage of fat cells was found to be significantly higher in siCITED2 treated group (25%) vs. siCONTROL treated group (7%). In addition, the total number of cells in both treatment groups were insignificantly different, indicating that expression knockdown of CITED2 during adipogenesis significantly promoted adipogenic differentiation efficiency of hMSCs, hence increasing the total number of mature fat cells. Furthermore, real time RT-PCR analysis demonstrated that the expression of CEBPα and PPARγ, two master regulators of adipogensis, were both dramatically increased in siCITED2 cells compare to siControl cells, suggesting that CITED2 normally acts as negative regulator of human adipogenesis by suppressing the expression of CEBPα and PPARγ.