useful for analyzing effects of drugs on metabolic syndrome such as obesity, diabetes and hypertension

White Adipocyte Culture Kit


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Background

White adipocytes, a type of subcutaneous adipocytes, play an important role in energy storage. White Adipocyte Culture Kit, V-1 (Rat) contains preadipocytes isolated from rat subcutaneous adipose tissues and culture medium that induces differentiation of precursor cells into mature adipocytes. The kit provides a convenient system for studying the mechanism of adipogenesis as well as for analyzing effects of drugs on metabolic syndrome such as obesity, diabetes and hypertension.

Feature and Advantages

• Search for obesity, diabetic, high blood pressure, and drug for arteriosclerosis
• Development of preventive food of lifestyle-related disease
• Functional test of antiobestic functional food
• Lipid metabolic experiment
• Thermal energy release experiment
• Function clarification of Brown Adipocyte
• Screening of new β3 agonist

Kit component

Product Name Size Quantity Storage Conditions Stability
Subcutaneous White Preadipocytes (SD Rat) 3.0 x 106 Cells 1 Liquid Nitrogen 2 years
-80°C Freezer 1 year
White Adipocyte Culture Medium
(Cat. No. PMC-SACMR-COS)
250 mL 1 -80°C Freezer 1 year
-20°C Freezer 6 months

Protocol (Cultured with 24-well plate)

1. Thaw Adipocyte Culture Medium in a 37°C water bath with gentle shaking.
2. Quickly place Common Precursor Cell vial in a 37°C water bath until the contents are thawed.
3. Transfer thawed cells into a 15 mL centrifuge tube containing 10 mL of Adipocyte Culture Medium. Mix gently and centrifuge at 4°C at 170 x g for 5 minutes.
4. After removing the supernatant, re-suspend cells in 10 mL of Adipocyte Culture Medium and centrifuge at 4°C at 170 x g for 5 minutes.
5. After removing the supernatant, re-suspend the cell pellet in 12.5 mL of Adipocyte Culture Medium.
6. Dispense 0.5 mL of cell suspension to each well of 24-well plate.
7. Incubate the plate at 37oC under 5% CO2, 100% humidity.
8. After 1 Day culture, add 0.5 ml of Adipocyte Culture Medium gently to each well of 24-well plate.
9. Change the medium every 2 days.  Be gentle not to disturb the cell layer.
    i. Approximately 3 days of culture, preadipocyte culture becomes confluent.
    ii. Approximately 4-5 days of culture, cells turn to mature adipocytes.
    iii. Approximately 7 days of culture, cells become hypertrophic.
    iv. Approximately 8 days of culture, start detaching from the bottom of the well.

Cellular morphology

References

> Hashimoto, T., Igarashi, J., Kosaka, H. Sphingosine Kinase is Induced in Mouse 3T3-L1 cells and Promotes Adipogenesis. J. Lipid Res. 50, 602-610 (2009)
> Takahashi, K., Yoshina, S., Masashi, M., Ito, W., Inoue, T., Shiwaku, H., Arai, H., Mitani, S., Okazawa, H. Nematode Homologue of PQBP1, a Mental Retardation Causative Gene, Is Involved in Lipid Metabolism. PLoS One. 4, e4104 (2009)
> Oguri, A., Tanaka, T., Iida, H., Meguro, K., Takano, H., Oonuma, H., Nishimura, S., Morita, T., Yamasoba, T., Nagai, R., Nakajima, T. Involvement of Cav3.1 T-type Calcium Channels in Cell Proliferation in Mouse Preadipocytes. Am. J. Physiol. Cell Physiol. 298, C1414-C1423 (2010)

Product List

Product Name Cat# Quantity Price

Subcutaneous Adipocyte Culture Kit V-1 DataSheet ,

PMC-SAC01-COS

1 SET ¥135,000
$1800

White Adipocyte Culture Medium DataSheet ,

PMC-SACMR-COS

250 ML ¥31,200
$416

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.



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