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View additional product information for PSC Cardiomyocyte Differentiation Kit - FAQs (A2921201)
35 product FAQs found
我们推荐用户在实验中一直使用H9或H7胚胎干细胞系作为对照。我们推荐用户针对难于分化的iPSC细胞系进行细胞密度的调整或延长诱导时间。
通过PSC心肌细胞分化试剂盒生成的心肌细胞经测试能够表达TNNT2，Nkx2.5，MYH6和α-辅肌动蛋白等关键标志物。心肌细胞免疫细胞化学试剂盒中包含了经验证过的抗体，能够检测用PSC心肌细胞分化试剂盒生成的培养物中TNNT2和Nkx2.5的表达。
分化的细胞可维持一个月或更长时间以进行长期研究。我们建议使用Gibco Geltrex基质用于长期培养。
分化得到的心肌细胞群体是由心房和心室细胞组成的混合体。随着时间的推移，培养物中心室细胞的占比会越来越大。
我们推荐使用EDTA进行PSCs的传代，在冻存前使用Gibco TrypLE酶分离心肌细胞。
每个试剂盒包含足够8块12孔板培养14天的体积。
我们推荐您使用Gibco Geltrex 无LDEV，经hESC验证的低生长因子基底膜基质（货号A1413301或A1413302）或在不含异源成份的应用中使用Gibco玻连蛋白（VTN-N）-截短型重组人源蛋白（货号A14700）。
我们不推荐冻存培养基。
是的。差异性的存在是正常的，发现某一细胞系未能有效分化也并不少见。在实验中包含一个对照细胞系——例如经验证能够良好分化的人ESC H1或H9细胞——可能会有帮助。
可以。心肌细胞维持培养基的单品的货号为A22920801。
分化后的细胞可维持一个月乃至更长，可供长期研究使用。我们推荐您搭配使用Gibco Geltrex基质用于长期培养。
通过PSC心肌细胞分化试剂盒生成的心肌细胞经测试能够表达TNNT2，Nkx2.5，MYH6和α-辅肌动蛋白等关键标志物。心肌细胞免疫细胞化学试剂盒中包含了经验证过的抗体，能够检测用PSC心肌细胞分化试剂盒生成的培养物中TNNT2和Nkx2.5的表达。
我们的研究结果显示分化操作起始阶段的细胞汇合度对于分化效率有着极其重要的影响。因此，我们推荐用户在开始分化操作时针对每一种PSC细胞系进行实验优化以确定最佳的细胞汇合度。相关的操作指南可在产品插页中找到。此外，我们还推荐在分化操作之前将细胞分散为单细胞而非小团块进行传代。在心肌细胞的分化实验中，将PSCs进行消化成单细胞的操作能够更好地估计接种密度和汇合度，从而在培养孔之间获得更稳定的实验结果以及总的说来能为一些难分化的细胞系提供更好的分化效果。
使用核型正常和确定表达多能性标志物的高品质人PSCs细胞（分化克隆的数目最低乃至无分化克隆）至关重要，这些细胞需以稳定接种间隔进行常规培养，维持健康的细胞外形直至开始心肌细胞的分化实验。此外，我们还建议不要使用传代次数超过100次的PSC细胞系。
通常情况下，得率是最初PSCs群体数目的100-200倍。以每块12孔板中含60万PSCs为例，您可获取6千5百万-7千万心肌细胞。
理想的平板是12孔板，因为它更能促进跳动合胞体的生成，方便收获和鉴定操作。
•心肌细胞分化培养基A：通过对BMP/ activin通路的激活和糖原激酶3的抑制，推动PSC向中胚层细胞转化。
•心肌细胞分化培养基B：通过抑制Wnt通路诱导心脏中胚层形成
•心肌细胞维持培养基：使心肌细胞成熟化
Yes. We have seen compatibility with the following differentiation kits provided by Thermo Fisher Scientific: PSC Cardiomyocyte Differentiation Kit (Cat. No. A2921201), PSC Definitive Endoderm Induction Kit (Cat. No. A3062601), PSC Neural Induction Medium (Cat. No. A1647801), and PSC Dopaminergic Neuron Differentiation Kit (Cat. No. A3147701).
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We recommend always using H9 or H7 ESC line as a control in your experiments. We recommend adjusting the cell density or extending the induction time for difficult-to-differentiate iPSC lines.
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Cardiomyocytes generated using PSC Cardiomyocyte Differentiation Kit have been tested for key markers such as TNNT2, Nkx2.5, MYH6, and Alpha-Actinin. The Cardiomyocyte Immunocytochemistry Kit (Cat. No. A25973) contains validated antibodies to measure TNNT2 and Nkx2.5 in cultures generated using PSC Cardiomyocyte Differentiation Kit.
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Differentiated cells can be maintained for a month or longer for long-term studies. We recommend the use of Gibco Geltrex Matrix for long-term cultures.
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The population of cardiomyocytes produced is a mix of atrial and ventricular cells. Over time, cultures become more ventricular.
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We recommend using EDTA for passaging PSCs and Gibco TrypLE Enzyme for dissociating cardiomyocytes prior to cryopreservation.
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Each kit contains enough volume for eight 12-well plates for 14 days of culture.
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We recommend using Gibco Geltrex LDEV-Free, hESC Qualified Reduced Growth Factor Basement Membrane Matrix (Cat. No. A1413301 or A1413302) or Gibco Vitronectin (VTN-N) Recombinant Human Protein, Truncated (Cat. No. A14700) for xeno-free applications.
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We do not recommend freezing the media.
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Yes. Variability is normal and it is not uncommon to find certain lines that will not differentiate as efficiently. Including a control line, such as the human ESC H1 or H9 cells, which have been shown to differentiate consistently well, may be helpful.
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Yes. Cardiomyocyte Maintenance Medium is sold separately as Cat. No. A22920801.
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Differentiated cells can be maintained for a month or longer for long-term studies. We recommend the use of Gibco Geltrex Matrix for long term cultures.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Cardiomyocytes generated using PSC Cardiomyocyte Differentiation Kit have been tested for key markers such as TNNT2, Nkx2.5, MYH6, and ?-Actinin. The Cardiomyocyte Immunocytochemistry Kit contains validated antibodies to measure TNNT2 and Nkx2.5 in cultures generated using PSC Cardiomyocyte Differentiation Kit.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Our studies have shown that the confluency of cells when starting differentiation can have a dramatic impact on efficiency of differentiation. Therefore, we recommend a range finding study to determine optimal confluency of each PSC line when starting differentiation. Guidance for this can be found in the product insert. Additionally, we recommend singularizing cells prior to differentiation rather than passaging in small clumps. Singularizing PSCs for differentiation to cardiomyocytes allows better seeding and confluence estimates, resulting in more consistent results well-to-well and overall better differentiation of difficult to differentiate lines.
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It is critical to use high quality human PSCs (with minimal or no differentiated colonies) that are karyotypically normal, confirmed to exhibit pluripotency markers, and are undergoing routine culture with regular subculture intervals and maintaining healthy morphology before starting cardiomyocyte differentiation. Additionally, we recommend that PSC line not be used past 100 passages.
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In general, yields will be 100-200 fold the starting population of PSCs. For example, from 0.6 million PSCs per 12-well plate, you can produce 65-70 million cardiomyocytes.
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The ideal plate format is 12-well due to the enhanced ability to create beating syncytium, easy harvesting, and characterization.
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Cardiomyocyte Differentiation Medium A: Pushes PSCs toward mesodermal commitment via BMP/activin pathway activation and glycogen kinase 3 inhibition
Cardiomyocyte Differentiation Medium B: Induces cardiac mesoderm via Wnt inhibition
Cardiomyocyte Maintenance Medium: Matures cardiomyocytes
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