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查看更多产品信息 Human Mammary Epithelial Cells (HMEC) - FAQs (A10565)
24 个常见问题解答
这一培养实验并不难。在标准的组织培养塑料器皿中，HMEC培养很容易以每平方厘米2.5 x 10E3个细胞开始，并通过胰酶/EDTA溶液进行传代操作。
单管细胞（500,000个活细胞）能够以以每平方厘米2500个活细胞的密度接种八个T-25培养瓶。应使用血细胞计数器对细胞进行精确计数，从而确保达到一个合适的接种密度。
这两种培养基均为HMEC的培养而专门设计。不过，内部研究显示在HMEC的生长速率和培养时间两个方面，HuMEC即用型培养基（Thermo Fisher Scientific货号12752-010）的性能均比M171/MEGS（Thermo Fisher Scientific货号M-171-500/S-015-5）更优。当使用非直接免疫细胞化学染色法（ICC）进行细胞特异性标志物的染色时，两种培养基之间并无差别。
在推荐的传代条件下，一次传代相当于4次左右的群体倍增。
通常在保质期内，HMEC的群体倍增时间为24-30小时。
HMEC细胞在复苏后可确保完成至少16次群体倍增。
是的。HMEC是从正常人体乳房缩小成形术所获得组织中分离得到的细胞。
对不同的原代细胞我们使用不同的培养基和添加剂。更多详细信息可查看此处 (https://www.thermofisher.com/us/en/home/life-science/cell-culture/primary-cell-culture.html).
通过每一培养物的接种密度和收获密度来计算累计群体倍增水平。
这一参数随着供体年龄、组织大小、位置及我们的设备条件而发生着很大变化。某些情况下，少量组织即可分离很多细胞；而某些情况下，大量组织只能取得少量细胞。通常情况下，我们可获得30-300管细胞。
是的，我们从组织中分离原代细胞需要使用酶进行消化。
原代细胞是从活体组织（例如活检材料）中直接获取，并在体外培养的细胞。由于这些细胞经历极少的群体倍增，因此比连续（肿瘤或人工永生化的）细胞系更能代表其来源组织的主要功能成分，使原代细胞成为体内状态更具代表性的模型。
通过使用不同种属来源的原代细胞，您可了解人类与临床前测试物种之间可能存在的差异。在开展体内研究之前，可使用小鼠或大鼠细胞摸索用量并减少临床前毒理实验所需的动物数量。人细胞可用于确定从动物模型外推所得人类数据的准确性。
No. HMEC cultures are easily initiated at 2.5 x 10E3 cells per cm2 in standard tissue culture plasticware and passaged using a trypsin/EDTA solution.
One vial (500,000 viable cells) can seed eight T-25 flasks at 2,500 viable cells per cm2. A hemocytometer should be used to accurately count cells and to help ensure an appropriate seeding density.
Both media are designed for culture of HMEC. However, in-house studies show HuMEC Ready Medium (Thermo Fisher Scientific Cat. No. 12752-010) outperforms M171/MEGS (Thermo Fisher Scientific Cat. No. M-171-500/S-015-5) in both growth rate and lifespan measurements of HMEC. There is no difference between media when staining for cell-specific markers using indirect ICC.
One passage is equivalent to approximately 4 population doublings when using the recommended subculture conditions.
HMEC usually have population doubling times of 24-30 hr during the guaranteed lifespan.
HMEC are guaranteed for a minimum of 16 population doublings after thaw.
Yes. HMEC are isolated from normal, human reduction mammoplasty tissue.
We use various media and supplements for the various primary cells. Details can be found here (https://www.thermofisher.com/us/en/home/life-science/cell-culture/primary-cell-culture.html).
Cumulative population doublings are calculated from the seeding density and harvesting density from each culture level.
It varies a lot depending on the age of donor, size of tissues, locations, and conditions arrived to our facility. Sometime a small amount of tissue gives us many cells and sometimes a large amount of tissue gives us a very small lot. Typically, we can get 30-300 vials.
Yes, our primary cells are isolated from tissue using enzymatic digestion.
Primary cells are cells taken directly from living tissue (e.g., biopsy material) and established for growth in vitro. These cells have undergone very few population doublings and are therefore more representative of the main functional component of the tissue from which they are derived in comparison to continuous (tumor or artificially immortalized) cell lines, making primary cells a more representative model for the in vivo state.
Primary cells from different species may be used, allowing you to highlight potential differences between humans and preclinical test species. Before in vivo studies, mouse or rat cells can be used to refine doses and reduce the number of animals required for preclinical toxicology. Human cells can be used to determine the accuracy of extrapolating human data from an animal model.