Journal of
Cell and Animal Biology

  • Abbreviation: J. Cell Anim. Biol.
  • Language: English
  • ISSN: 1996-0867
  • DOI: 10.5897/JCAB
  • Start Year: 2007
  • Published Articles: 260

Full Length Research Paper

Growth arrest induction of 3T3-L1 preadipocytes by serum starvation and their differentiation by the hormonal adipogenic cocktail

Zhenhui Cao1#, Hui Yang2#, Lingfu Kong1, Dahai Gu2, Zhanlong He3, Zhiqiang Xu2, Junjing Jia4, Changrong Ge4* and Qiuye Lin1
1Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, P. R. China. 2College of Food Science and Technology, Yunnan Agricultural University, Kunming, P. R. China. 3Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming, P. R. China. 4Key Laboratory of Animal Nutrition and Feed Science, Kunming, P. R. China.
Email: [email protected]

  •  Accepted: 22 February 2012
  •  Published: 31 March 2012

Abstract

The current methods for differentiation of 3T3-L1 preadipocytes start with growth arrest by contact inhibition. However, on day 6, 3T3-L1 preadipocytes reliably detach and differentiation could no longer proceed. In the present study, we used serum starvation to induce growth arrest in 3T3-L1 preadipocytes and investigated their differentiation by the modified hormonal adipogenic cocktail.  About 85% of 3T3-L1 cells were in G0/G1 stage under serum deprivation. There were no significant difference between the percentage of cells in G0/G1 phase under these serum deprivation condition and that of post-confluent cells. Growth assays indicated that cells under serum starvation condition grew slow, two-days later than growing and post-confluent cells. We determined that 0.5% FBS for 48 h was the optimal condition to arrest 3T3-L1 cells at the G0/G1 phase. The grow-arrested 3T3-L1 preadipocytes were induced to differentiation by hormonal adipogenic cocktail. After 19 days of differentiation, over 90% 3T3-L1 cells exhibited adipocyte morphology with ring-like lipid droplet in cytoplasm. The mRNA levels of critical transcriptional factor CCAAT/enhancer binding proteins-α and peroxisome proliferator-activated receptor-γ were determined to validate the differentiation. These results indicated that serum starvation effectively arrested the growth of 3T3-L1 preadipocytes and prevented cells from detachemnt caused by contact inhibition at confluence. Furthermore, growth arrested 3T3-L1 preadipocytes by serum starvation successfully differentiated into adipocytes by the hormonal adipogenic cocktail.

 

Key words: Cell cycle, detachment, differentiation, growth arrest, preadipocytes, serum starvation,