| Neurosphere Co-culture Assay
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Author:
Date:
2016-08-05
[Abstract] The hippocampal niche is one of two areas in the brain where stem cells reside. In this neurogenic niche, stem cells can be found in close proximity to astrocytes and in contact with microvessels consisting of pericytes and endothelial cells. To study the regulatory interplay of this complex niche network in a simplified in vitro model, we established a co-culture system. We investigate the formation of neurosphere under different co-culture conditions by using primary niche cells. Here, we describe the isolation procedure for primary niche cells culture of astrocytes, endothelial cells and pericytes/smooth muscle cells from mouse brain. These niche cells are co-cultured (by hanging inserts) with freshly isolated stem and precursor cells from the adult hippocampus to study the ...
[摘要] 真菌形态发生需要细胞壁的修饰和可塑性，这意味着其组分（包括壳多糖和葡聚糖）的合成和重塑。因此几丁质酶和葡聚糖酶活性对于细胞壁生物发生和细胞分裂是至关重要的。几丁质酶活性的定量可能有助于鉴定可能负面影响一些丝状真菌如产生细胞内和分泌型几丁质酶的尖孢镰刀菌的生长和形态发生的结构缺陷。基于它们对壳多糖底物的酶作用将壳聚糖分解酶分类。内酯酶被定义为催化几丁质链中内部点的随机裂解的酶。外切球蛋白酶催化乙酰壳寡糖或N-乙酰葡糖胺从几丁质的非还原端逐步释放，因此分别称为壳聚糖酶和β-N-乙酰氨基葡糖苷酶。在这里，我们描述了一种简单的方法来轻易地纯化几丁质酶，以便比较不同的F的内切几丁质酶活性和外切几丁质酶活性。尖孢镰孢菌株。该方案可适用于任何真菌物种。...
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| Trichome Isolation and Integrity Test from Brassica villosa and Other Species
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Author:
Date:
2014-12-20
[Abstract] The outward growths from one or more epidermal cells are well known as trichomes (plant hair cells) (Levin, 1973; Mathur, 2006). Preparation of pure intact non-glandular trichomes from trichome-rich Brassica villosa depended on rendering the trichomes sufficiently stiff to dislodge them from the leaves in an undamaged state, which can be further used for structural, transcriptome, genome and biochemical or chemical analysis. Dislodging the flexible trichomes from Brassica villosa (B. villosa) with a paintbrush [as in Zhang and Oppenheimer (2004)] proved too gentle, and scraping trichomes off the leaves with a razor blade [as in Zhang and Oppenheimer (2004)] resulted in trichome cell disruption. Aziz et al. (2005) reported isolating alfalfa glandular ...
[摘要] 来自一种或多种表皮细胞的向外生长是众所周知的毛状体（植物毛细胞）（Levin，1973; Mathur，2006）。从富含毛藓的芸苔制备纯的完整无腺毛状体依赖于使毛状体足够坚硬，以便在未受损的状态下将它们从叶子上除去，这可以进一步用于结构，转录组，基因组和生化或化学分析。用油漆刷（如Zhang和Oppenheimer（2004）中所述）从芸苔属villosa（ B。villosa ）中移出柔性毛状体证明太柔和，剃刀刀片[如Zhang和Oppenheimer（2004）]导致毛状体细胞破裂。 Aziz等人（2005）报道了通过在液氮中剪切来分离苜蓿腺毛状体（N 2 sub）。在本研究中，使用类似的方法。通过用液体N 2处理组织以硬化柔性毛状体，接着用共同的涡旋混合器剪切1分钟的剪切力以从其叶床上除去毛状体来分离非腺体叶毛状体（Nayidu et al。 em> et al。，2014）。高达〜20％的毛状体从叶表面除去，并且大多数未破裂（通过台盼蓝没有染色证实;图1）。然后通过使组织/毛/水混合物通过筛子来纯化毛状体。然而，如果将叶组织搅拌较长时间，则在浸入液体N 2后叶组织也变得非常脆，并且分裂成非常小的碎片。因此，N ...
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