Effect of Leukemia Inhibitory Factor and Forskolin on Establishment of Rat Embryonic Stem Cell Lines

This study was designed to investigate whether supplementation of 2i medium with leukemia inhibitory factor (LIF) and/or forskolin would support establishment of germline-competent rat embryonic stem (ES) cell lines. Due to the higher likelihood of outgrowth rates, supplementation of forskolin with or without LIF contributed to the higher establishment efficiency of ES cell lines in the WDB strain. Germline transmission competency of the chimeric rats was not influenced by the profile of ES cell lines until their establishment. When the LIF/forskolin-supplemented 2i medium was used, the rat strain used as the blastocyst donor, such as the WI strain, was a possible factor negatively influencing the establishment efficiency of ES cell lines. Once ES cell lines were established, all lines were found to be germline-competent by a progeny test in chimeric rats. In conclusion, both LIF and forskolin are not essential but can play a beneficial role in the establishment of “genuine” rat ES cell lines.     

Forskolin和促性腺激素对猪卵母细胞体外成熟的影响

利用已建立的猪卵母细胞体外无血清培养模型,研究了猪卵丘-卵母细胞复合体(CEO)在模拟生理环境的培养条件下Forskolin和促性腺激素对猪卵母细胞成熟的作用.在观察卵母细胞减数分裂恢复(GVBD)后的实验结果表明(1)cAMP产生的激动剂凡Forkolin(3μmol@L-1)处理CEO 20h可以显著协同HX抑制卵母细胞成熟;处理40h后,Forskolin(3μmol@L-1)的抑制作用被卵丘细胞分泌的某种物质克服;(2)Forskolin(3μmol L.)短期(30,60,120,240min)处理后可以诱导卵丘细胞分泌某种促进卵母细胞成熟的物质.(3)FSH(50～200U/L)能够诱导卵丘细胞分泌某种物质促进卵母细胞成熟;(4)50U/L的FSH预处理CEO结果与Forskolin结果相同,提示FSH诱导的卵母细胞成熟是通过cAMP依赖性的通路进行的.     

Human mesenchymal stem cells differentiate into neuron-like cells by increase in intracellular cyclic AMP

AIM: To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells by the increase in intracellular cyclic AMP. METHODS: hMSC were separated from human marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with Forskolin and 3-isobutyl-1-methyl-xanthine (IBMX). Neuron-specific enolase(NSE), neurofilament(NF), glial fibrillary acaidic protein(GFAP) were detected by immunohistochemistry. RESULTS: hMSC were expanded as undifferentiated cells in culture for more than10passages. Forskolin/IBMX can induce hMSC to exhibit a neuronal phenotype, expressing NSE and NF-M in 5 hours. But the neuron-like cells didn't express the glial astrocyte marker GFAP. CONCLUSION: hMSC can be induced to differentiate into neurons by increase in the intracellular cAMP.     

Zinc is involved in regulation of secretion from intestinal epithelium in weaned piglets

The objective was to study the effect of serosal zinc (Zn) on intestinal chloride (Cl⁻) secretion in vitro by the Ussing chamber technique. The secretagogues used to stimulate Cl⁻ secretion were serotonin (5-HT), vasoactive intestinal polypeptide (VIP) and forskolin (FSK). In addition correlations between organ and plasma Zn levels vs. the responses to 5-HT were studied. The results revealed an attenuating effect of serosal Zn on the secretory response to 5-HT, VIP and FSK. Furthermore, negative correlations between secretory responses to 5-HT vs. ADG and plasma Zn concentrations were found, whereas the responses to 5-HT did not correlate to the Zn concentration in liver or intestinal mucosa. It is suggested, that dietary Zn reduces diarrhoea directly through a regulatory role of serosal Zn on Cl⁻ secretion and indirectly by improving the nutritional status, which may stabilize the function of the intestinal epithelium.