Differentiation and function of rat adipocyte precursor cells in primary culture.
Differentiation and function of rat adipocyte precursor cells in primary culture.
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发表时间：
1980-08
影响因子：
6.5
通讯作者：
P. Björntorp;M. Karlsson;P. Pettersson;G. Sypniewska
中科院分区：
文献类型：
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作者：
P. Björntorp;M. Karlsson;P. Pettersson;G. Sypniewska
从小鼠附睾脂肪垫中纯化脂肪细胞前体细胞并进行原代培养研究。利用琼脂糖饲喂层，采用底物和辅因子对完全脂肪细胞转化不限速的培养体系。通过添加含有甲基纤维素的粘性培养基层，可以防止细胞从培养皿中脱离。该系统允许对形成的脂肪细胞进行定量和明确的表征，脂肪细胞被定义为直径为20微米的脂滴。细胞可传代培养，但逐渐丧失转化脂肪细胞的能力。捐献者的年龄抑制了脂肪细胞的转化，然而，这种转化似乎从未完全停止过。前列腺素E(1)和F（2alpha）在生理浓度范围内无明确作用，吲哚美辛可能有弱抑制作用。胰岛素、肝素和异丁基甲基黄嘌呤增加脂肪细胞的形成。观察脂肪细胞特征功能随时间的变化。在分离的前体细胞中，脂蛋白脂肪酶活性在培养前很低，但在融合培养中发展起来，并在几天内提高了1000倍。在这个峰值脂蛋白脂肪酶活性比来自同一供体动物的成熟脂肪细胞高50倍。葡萄糖合成甘油三酯达到平行峰值，但从未达到成熟脂肪细胞的值，合成的脂肪酸很少。激素敏感的甘油释放在汇合时发展并达到成熟脂肪细胞的活性水平。这项研究和以前的工作已经表明环化酶系统在前体细胞脂肪细胞的发育中起作用。二丁基环AMP引起脂蛋白脂肪酶活性和脂肪细胞转化的增强。在悬浮培养基中，核苷酸引起抑制。这些结果与核苷酸的影响相一致，不直接影响脂蛋白脂肪酶和细胞测定，而是通过与细胞间相互作用相关的汇合发生的事件。与先前描述的经过脂肪转化的已建立细胞系的细胞（3T3细胞）相比，本研究中描述的细胞，与脂肪细胞一样，在外源性脂质来源（脂蛋白脂肪酶活性）的脂肪酸结合途径中，比在重新合成的途径中表现出更多的代谢活性。此外，脂肪细胞形成的年龄依赖性和部位依赖性等宿主因素也可以被追踪。生理刺激如胰岛素、脂质底物和肝素对脂肪细胞形成有影响。由此得出结论，这种细胞制备在脂肪细胞增殖调控的研究中具有潜在的生理意义。-Björntorp， P.， M. Karlsson， P. Pettersson和G. Sypniewska。原代培养大鼠脂肪前体细胞的分化及功能。
Precursor cells to adipocytes were purified from the epididymal fat pads of small rats and studied in primary culture. A culture system in which substrate and cofactors were not rate-limiting for complete adipocyte conversion was used by utilizing an agarose feeding-layer. Detachment of cells from the culture dish was prevented by addition of a viscous layer of culture medium, containing methyl cellulose. This system allowed quantitation and definite characterization of formed adipocytes, defined as cells accumulating a lipid droplet >20 micro m in diameter. The cells could be subcultured but then gradually lost their adipocyte conversion ability. Age of the donor depressed the adipocyte conversion which, however, never seemed to stop completely. Prostaglandin E(1) and F(2alpha) had no definite effect in the physiological concentration range while indomethachin possibly had a weak inhibitory effect. Insulin, heparin, and isobutylmethylxanthine increased adipocyte formation. Development of characteristic adipocyte functions with time was examined. Lipoprotein lipase activity was very low in the isolated precursor cells before culture, but developed in culture at confluence and was a thousand-fold higher within a few days. At this peak lipoprotein lipase activity was 50-fold higher than in mature adipocytes from the same donor animal. Triglyceride synthesis from glucose peaked in parallel but never reached the value of mature adipocytes and very little fatty acid was synthesized. Hormone-sensitive glycerol release developed at confluence and reached the level of activity of mature adipocytes. This study and previous work have indicated a role for the cyclase system in the development of adipocytes from precursor cells. Dibutyrylcyclic AMP caused an enhancement of lipoprotein lipase activity and adipocyte conversion. In suspension media, the nucleotide caused inhibition. These results are compatible with an effect of the nucleotide, not directly on lipoprotein lipase and cell determination, but via events taking place at confluence associated with cell to cell interactions. In comparison with previously described cells from an established cell line which undergo adipose conversion (3T3 cells), the cells described in the present work, like adipocytes, showed more metabolic activity in pathways for fatty acid incorporation from exogenous lipid sources (lipoprotein lipase activity) than from de novo synthesis. Furthermore, host-factors could be followed such as in the age- and site-dependence of adipocyte formation. Physiological stimuli such as insulin, lipid substrate, and heparin had effects on adipocyte formation. It was therefore concluded that this cell preparation has a potential of yielding information of physiological significance in studies of the regulation of adipocyte multiplication.-Björntorp, P., M. Karlsson, P. Pettersson, and G. Sypniewska. Differentiation and function of rat adipocyte precursor cells in primary culture.