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2019 | 25 |

Tytuł artykułu

Seeding cell number required for optimal lipid accumulation during adipocyte differentiation using 3T3-L1 cell line

Treść / Zawartość

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
Obesity is one of the major causes of metabolic diseases such as diabetes and heart attack, and, hence, can lead to low quality of life. Elaborating adipocyte differentiation is very crucial for formulating the treatment and prevention of obesity. The objective of this study is to investigate the seeding cell number required to obtain optimum lipid accumulation during adipocyte differentiation using the 3T3-L1 cell line. Two sets of 5.48×104 (for Day 0 and Day 8 of differentiation), of 10.96×104 (for Day 8) and of 21.92×104 (for Day 8) of 3T3-L1 cells were seeded in each wells of a 12-well plate. Isobuthylmethylxanthine (IBMX), Dexamethasone, and Insulin-containing differentiation cocktails was added into the medium at Day 0 for 48 hours. The medium was changed every two days. Day 0 and Day 8 samples were then stained using Oil Red O and were examined under the microscope to observe the lipid droplets (red-coloured). The lipid droplets were quantified by measuring the absorbance at wavelength of 550 nm. In the study, seeding the number of 10.96×104 cells produced very significantly higher lipid accumulation, as compared with seeding the number of 5.48×104 cells. However, doubling the seeding number into 21.92×104 cells did not increase the lipid droplets significantly. This study found that the optimum seeding number to obtain the maximum lipid droplets during 3T3-L1 adipocyte differentiation was 10.96×104 cells.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

25

Opis fizyczny

p.220-226,fig.,ref.

Twórcy

  • Division of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Jl. Raya Bandung Sumedang, KM.21, Sumedang, Indonesia
  • Research Center for Medical Genetics, Faculty of Medicine, Universitas Padjadjaran, Jl. Eyckman No. 38, Bandung, Indonesia
  • Faculty of Medicine, Universitas Padjadjaran, Jl. Raya Bandung Sumedang, KM.21, Sumedang, Indonesia
autor
  • Department of Psychiatry, Faculty of Medicine, Universitas Padjadjaran / Dr. Hasan Sadikin General Hospital, Jl. Pasteur No.38, Bandung, Indonesia
  • Cell Culture Laboratory, Faculty of Medicine, Universitas Padjadjaran, Jl. Eyckman No. 38, Bandung, Indonesia
autor
  • Cell Culture Laboratory, Faculty of Medicine, Universitas Padjadjaran, Jl. Eyckman No. 38, Bandung, Indonesia
autor
  • Division of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Jl. Raya Bandung Sumedang, KM.21, Sumedang, Indonesia
autor
  • Division of Biochemistry and Molecular Biology, Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Jl. Raya Bandung Sumedang, KM.21, Sumedang, Indonesia

Bibliografia

  • [1] Rachmi CN, Li M, Alison Baur L. Overweight and obesity in Indonesia: prevalence and risk factors - a literature review. Public Health 2017 Jun, 147: 20-29
  • [2] Arundhana AI, Utami AP, Dwi Muqni A, Thalavera MT. Regional differences in obesity prevalence and associated factors among adults. Mal J Nutr Vol. 24(2) (2018) 193-201
  • [3] Ruiz-Ojeda FJ, Rupérez AI, Gomez-Llorente C, Gil A, Aguilera CM. Cell Models and Their Application for Studying Adipogenic Differentiation in Relation to Obesity: A Review. Int J Mol Sci . 2016 Jun 30, 17(7).
  • [4] Suemori H, Yasuchika K, Hasegawa K, Fujioka T, Tsuneyoshi N, Nakatsuji N. Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage. Biochem Biophys Res Commun 2006 Jul; 345(3): 926–32
  • [5] Marques ES, Leite TH, Azeredo CM, Cunha DB, Verly Júnior E. Effective strategies for prevention, control, and treatment of obesity in primary health care setting for adolescents, adults, and elderly people. Medicine (Baltimore) 2018 Jun; 97(22): e10925
  • [6] Sung J-H, Chon J-W, Lee M-A, Park J-K, Woo J-T, Park YK. The anti-obesity effect of Lethariella cladonioides in 3T3-L1 cells and obese mice. Nutr Res Pract . 2011 Dec; 5(6): 503–10
  • [7] Zebisch K, Voigt V, Wabitsch M, Brandsch M. Protocol for effective differentiation of 3T3-L1 cells to adipocytes. Anal Biochem. 2012 Jun 1, 425(1): 88–90
  • [8] Pu Y, Veiga-Lopez A. PPARγ agonist through the terminal differentiation phase is essential for adipogenic differentiation of fetal ovine preadipocytes. Cell Mol Biol Lett. 2017 Dec 23; 22(1): 6
  • [9] Hausman GJ, Basu U, Wei S, Hausman DB, Dodson MV. Preadipocyte and Adipose Tissue Differentiation in Meat Animals: Influence of Species and Anatomical Location. Annu Rev Anim Biosci 2014 Feb, 2(1): 323–351
  • [10] Kaur G, Dufour JM. Cell lines: Valuable tools or useless artifacts. Spermatogenesis 2012 Jan 1, 2(1): 1–5.
  • [11] Chavey C., Lagarrigue S., Annicotte JS., Fajas L. (2013) Emerging Roles of Cell Cycle Regulators in Adipocyte Metabolism. In: Bastard JP., Fève B. (eds) Physiology and Physiopathology of Adipose Tissue. Springer, Paris
  • [12] Suemori H, Yasuchika K, Hasegawa K, Fujioka T, Tsuneyoshi N, Nakatsuji N. Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage. Biochem Biophys Res Commun. 2006 Jul; 345(3): 926–32
  • [13] Kawai M, Namba N, Mushiake S, Etani Y, Nishimura R, Makishima M, et al. Growth hormone stimulates adipogenesis of 3T3-L1 cells through activation of the Stat5A/5B-PPARgamma pathway. J Mol Endocrinol 2007 Feb 1, 38(1–2): 19–34
  • [14] Cadena-Herrera D, Esparza-De Lara JE, Ramírez-Ibañez ND, López-Morales CA, Pérez NO, Flores-Ortiz LF, et al. Validation of three viable-cell counting methods: Manual, semi-automated, and automated. Biotechnol Rep (Amst). 2015 Apr 18; 7: 9-16. doi: 10.1016/j.btre.2015.04.004
  • [15] Hudu SA, Alshrari AS, Syahida A, Sekawi Z. Cell Culture, Technology: Enhancing the Culture of Diagnosing Human Diseases. J Clin Diagn Res 2016 Mar; 10(3): DE01-5.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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