ABSTRACT
Silybum marianum [L.] Gaertn is a kind of medicinal plants. Silymarin is a derivate substance from the fruits of milk thistle plant which is consists of a large number of flavonolignans. Cell cultures derived from this species could be an alternative for production of flavonolignans. Elicitors cause enhancement in metabolite production by effecting on key enzymes in secondary metabolites pathways. In this study various level of Yeast extract in 5 different exposure time have been used as an biotic elicitor in order to evaluation the effect of Yeast extract on silymarin production and cell growth and nomination the best time and best consistence of the elicitor. In this study various level of Yeast extract in 5 different exposure time have been used as an biotic elicitor in order to evaluation the effect of Yeast extract on silymarin production and cell growth and nomination the best time and best consistence of the elicitor. In this work after preparation cell suspension culture of S. marianum, the effects of various level of yeast extract [1, 2. 4, 6 and 8 mg/ 50 ml culture] in 6 different exposure time [12,24, 48, 72, 144 and 216 h] on flavonolignans production by High Performance Liquid Chromatography [HPLC] have been studied. Determination and quantification of flavonolignans showed that cell suspension cultures of S.marianum were consists of a large number of flavonolignans including silychristin, silydianin, silybin, isosilybin and taxifolin. The results showed that Yeast extract cause improvement in silymarin content in the media treated with 6 [mg/ 50 ml culture] Yeast extract at the end of 72h which was 5- fold to compare the control and the maximum cell Dry weight was 5.82 g in the media treated with 4 mg/ 50 ml culture Yeast extract at the end of 24h. In this experiment it has been observed that cell suspension culture of S. marianum are susceptible to elicitation by Yeast extract and it can be extremely useful in increasing productivities in cell suspension culture of Milk thistle plant
Subject(s)
Silymarin , Flavonolignans , Cells, Cultured , YeastsABSTRACT
Introduction: Extra cellular matrix [ECM] as an important component of cellular microenvironment has a key role in maintaining the differentiated state of cells. Effects of ECM on morphologic differentiation of epithelial cells including those from uterus and oviduct has been shown in past studies in which cellular and hormonal factors have been used in addition to ECM to maintain epithelial cell differentiation. Not much attention has been paid, in these studies; about the ultra structure of cultured cells specially those from oviduct
Objective: The purpose of present study is to cultivate the human uterine and oviduct epithelial cells under the same microenvironment [ECM Gel and DMEM/Ham's F12 medium] and to observe and compare ultra structural characteristics of the cultured cells by transmission electron microscopy [TEM]
Materials and Methods: For this purpose, uterine and oviduct tissue were obtained from patients undergoing total hysterectomy in Emam Khomeini Hospital. Epithelial cells, after being isolated, were cultured on plastic surfaces and the epithelial nature of the cells was confirmed using immunocytochemistry. Cells with epithelial nature were trypsinized and cultured on ECM gel. At the end ultra structure of cells in parallel with tissue were prepared for TEM
Results: Our results showed that the plastic cultured cells have no signs of differentiation and appeared as elongated spindle cell in sections, whereas those cultured on ECM gel had highly differentiated structure and observed as columnar in shape. In this term they were very similar to epithelial cells from tissue fragment. Epithelial cells of oviduct, cultured on ECM gel, were noticed ultra structurally very similar to that from uterus. The main structural difference existed in vivo state [the presence of abundance cilia on apical surface of oviduct epithelial cells] were not observed in vitro
Conclusion: As a conclusion, it seems that ECM gel by itself is enough to induce morphologic differentiation and structural polarization of epithelial cells. Ultra structurally different cells grows and acquires the same structure when being cultured under the same microenvironment
ABSTRACT
Introduction: Extracellular matrix [ECM] components significantly influence the growth characteristics of cell, development of spontaneous contractile activity, and morphologic differentiation. The present study evaluates the effect of ECM on genotypic and physiologic characteristics of cardiomyocytes derived from embryonic stem [ES] cells
Material and Methods: Embryoid bodies derived from mouse ES cells line Royan B1 [C57BL/6] were cultured on commercial ECM [matrigel], a cardiac fibroblast-derived extracellular matrix [cardiogel], and control media [without ECM] for 21 days. The growth characteristics of cardiomyocytes were assessed by immunocytochemistery, RT-PCR, and chronotropic drugs
Results: The beating frequency of cardiomyocytes cultured on cardiogel increasedin comparison with cardiomyocytes cultured on matrigel and/or control during days 7-9 [p<0.05] The spontaneously beating cells expressed markers characteristic of cardiomyocytes including alpha-actinin, desmin, tropnin-I, sarcomeric myosin heavy chain [MHC], pan-cadherin, connexin 43, cardiac alpha -MHC, cardiacbeta-MHC, myosin light chain isoform-2V, and atrial natriuretic factor. In addition, spontaneousness of cardiomyocytes on both ECMs was enhanced by treatment of cells with isopernaline, while reduced more on matrigel and carbacol when compared with control and cardiogel. However, the change in beating was similar in all groups upon treatment with phenylephrine, propranolol and for Bay-K
Conclusions: We conclude that the ECM influences ES-derived cardiomyocytes, in terms of chronotropic characteristics