Hostname: page-component-78c5997874-dh8gc Total loading time: 0 Render date: 2024-11-05T01:11:04.234Z Has data issue: false hasContentIssue false

Isolation and in vitro culture of intramuscular pre-adipocytes from Luxi adult Yellow cattle

Published online by Cambridge University Press:  13 February 2008

Wan Rong
Affiliation:
The State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, and Beef Cattle Research Center, China Agricultural University, Beijing 100094, China
Ding Jian
Affiliation:
The State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, and Beef Cattle Research Center, China Agricultural University, Beijing 100094, China
Zhou Zhen-Ming
Affiliation:
The State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, and Beef Cattle Research Center, China Agricultural University, Beijing 100094, China
Ren Li-Ping
Affiliation:
The State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, and Beef Cattle Research Center, China Agricultural University, Beijing 100094, China
Meng Qing-Xiang*
Affiliation:
The State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, and Beef Cattle Research Center, China Agricultural University, Beijing 100094, China
*
*Corresponding author. E-mail: [email protected]

Abstract

Three Luxi adult Yellow steers were used to isolate and culture intramuscular pre-adipocytes in vitro as well as to examine factors influencing their proliferation and differentiation. The intramuscular pre-adipocytes were taken from adipose tissues within muscles between the sixth and seventh rib and cultured after digestion with collagenase I. The results showed that the separated cell populations were highly homogeneous, proliferative and doubled within 62 h. When the confluent pre-adipocytes were treated with 10 μg/ml insulin and 0.25 μmol/l dexamethasone, small lipid droplets appeared on day 2 and the number of lipid droplets rapidly increased around the nuclei on day 6. Their dynamic morphological changes, growth curve, Oil Red O staining, and reaction to insulin and dexamethasone all verified their pre-adipocyte identity. Under controlled conditions, the intramuscular pre-adipocytes resumed proliferating and differentiating in vitro. Interestingly, the proportion of cultured diploid pre-adipocytes reached more than 90% after six repeated cultures. This study confirms the existence of functionally active pre-adipocytes within the muscles of Chinese adult local breed cattle. These cell strains are a potentially useful model for understanding further the mechanism of intramuscular adipose deposition in tissues, in order to improve beef quality based on Chinese local breed beef cattle.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

First published in Journal of Agricultural Biotechnology 2007, 15(3): 419–423

References

Albrecht, E, Teuscher, F, Ender, K and Wegner, J (2006) Growth- and breed-related changes of marbling characteristics in cattle. Journal of Animal Science 84: 10671075.CrossRefGoogle ScholarPubMed
Aso, H, Abe, H, Nakajima, N, et al. (1995) A preadipocyte clonal line from bovine intramuscular adipose tissue: nonexpression of GLUT-4 protein during adipocyte differentiation. Biochemical and Biophysical Research Communications 213(2): 369375.CrossRefGoogle ScholarPubMed
Carraro, R, Li, ZH, Johnson, JE Jr and Gregerman, RI (1991) Islets of preadipocytes highly committed to differentiation in cultures of adherent rat adipocytes. Cell and Tissue Research 264(2): 243251.CrossRefGoogle ScholarPubMed
Nishimura, T, Hattori, A and Takahashi, K (1999) Structural changes in intramuscular connective tissue during the fattening of Japanese black cattle: effect of marbling on beef tenderization. Journal of Animal Science 77: 93104.CrossRefGoogle ScholarPubMed
Ohyama, M, Matsuda, K, Torii, S, et al. (1998) The interaction between vitamin A and thiazolidinedione on bovine adipocyte differentiation in primary culture. Journal of Animal Science 76: 6165.CrossRefGoogle ScholarPubMed
Qu, CQ, Zhang, GH, Chen, FF, Zhao, LL and Yang, GS (2005) Primary culture of porcine preadipocyte. Journal of Agricultural Biotechnology 13(5): 649653 (in Chinese with English abstract).Google Scholar
Ramirez-Zacarias, JL, Castro-Munozledo, F and Kuri-Harcuch, W (1992) Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red O. Histochemistry 97: 493497.CrossRefGoogle ScholarPubMed
Sato, K, Nakanishi, N and Mitsumoto, M (1996) Culture conditions supporting adipose conversion of stromal-vascular cells from bovine intramuscular adipose tissues. Journal of Veterinary Medical Science 58(11): 10731078.CrossRefGoogle ScholarPubMed
SiTu, ZQ (2001) Cell Culture. Xi'an: World Publishing Co, Ltd (in Chinese).Google Scholar
Sun, C (2001) Regulation of mice preadipocytes' proliferation and differentiation by ECM components and cAMP. PhD thesis, Northwest A&F University (in Chinese with English abstract).Google Scholar
Torii, S, Kawada, T, Matsuda, K, Matsui, T, Ishihara, T and Yano, H (1998) Thiazolidinedione induces the adipose differentiation of fibroblast-like cells resident within bovine skeletal muscle. Cell Biology International 22(6): 421427.CrossRefGoogle ScholarPubMed
Van, RLR, Bayliss, CE and Roncari, DAK (1976) Cytological and enzymological characterization of human adult adipocyte precursors in culture. Journal of Clinical Investigation 58: 699704.CrossRefGoogle ScholarPubMed
Wang, ZC, Liu, JZ, Li, Y, Yang, DZ and Kuang, JQ (2001) Primary culture of human preadipocyte. Academic Journal of Sun Yat-Sen University of Medical Science 22(6): 443447 (in Chinese with English abstract).Google Scholar
Zhang, JB and Xu, CS (2004) Culture of Animal Cells. Beijing: Science Publishing Co. Ltd (in Chinese).Google Scholar
Zhou, GH, Liu, L, Sun, BG, et al. (2001) The methods and standard of the beef grading system. The Meat Industry 241(6): 4151 (in Chinese with English abstract).Google Scholar