产品号 #100-1524_C
从人类多能干细胞稳健生成脊髓类器官的细胞培养基试剂盒
若您需要咨询产品或有任何技术问题,请通过官方电话 400 885 9050 或邮箱 info.cn@stemcell.com 与我们联系
从人类多能干细胞稳健生成脊髓类器官的细胞培养基试剂盒
cGMP, enzyme-free cell dissociation reagent
Cell culture medium kit for long-term maintenance and maturation of patterned neural organoids
Microwell culture plates for easy and reproducible production of embryoid bodies and spheroids
Compatible antibodies for purity assessment of isolated cells
使用 STEMdiff™ 脊髓类器官分化试剂盒,可无需基质嵌入,稳健地从人多能干细胞生成三维、模式化的脊髓类器官培养物。配合AggreWell™800使用时,该无血清细胞培养体系可防止类器官融合,每套试剂盒最多可生成 300 个类器官,从而实现更高精度的统计重复和更详细的纵向研究。
该试剂盒基于 Dr. Sergiu Paşca的实验方法开发(Andersen J 等,Cell,2020;Yoon SJ 等,Nature Methods,2018),所生成的脊髓类器官是三维体外模型,具有人类颈部脊髓发育阶段所代表的细胞组成和结构特征。它们在不同细胞系之间具有一致的大小和可复制的形态,为在功能性三维系统中研究运动神经元提供了可靠模型。
使用该试剂盒生成的类器官也可以作为AssemBloids™共培养,以研究脑区相互作用((Andersen J et al. Cell, 2020)。对于较长时间(> 50 天)的类器官培养,可以使用STEMdiff™神经类器官维持试剂盒进行维持,以支持类器官的长期培养存活,从而进行预测性分析、高通量表型筛选和神经毒性分析。
亚型
专用培养基
细胞类型
神经细胞,PSC衍生,神经干/祖细胞,多能干细胞
应用
细胞培养,鉴定,分化,功能学筛选,免疫荧光,类器官培养,表型鉴定,球状体培养
品牌
STEMdiff
研究领域
疾病建模,药物发现和毒理检测,神经科学,类器官
制剂类别
无血清
Figure 1. Schematic for the STEMdiff™ Spinal Cord Organoid Differentiation Kit
hPSC-derived spinal cord organoids can be generated in 43 days with the STEMdiff™ Spinal Cord Organoid Differentiation Kit. Organoids are initially formed in AggreWell™800 plates. After 6 days, the organoids are transferred and cultured in suspension, allowing growth and subsequent patterning to the spinal cord. Long-term maintenance and further maturation of spinal cord organoids can be achieved using STEMdiff™ Neural Organoid Maintenance Kit, see the Product Information Sheet (PIS) for details. hPSC = human pluripotent stem cell
Figure 2. STEMdiff™ Spinal Cord Organoids Express Expected Progenitor Markers
Spinal cord organoids were generated from ES cell line H1 with STEMdiff™ Spinal Cord Organoid Differentiation Kit and express expected progenitor markers via immunostaining. (A) At Day 19, the organoids express elevated levels of the cortical progenitor marker PAX6 in the interior and exterior regions of the organoid as well as (B) motor neuron progenitor marker OLIG2 in the exterior region of the organoid. ES = embryonic stem
Figure 3. STEMdiff™ Spinal Cord Organoids Express Expected Markers
Spinal cord organoids were generated from hPSCs with STEMdiff™ Spinal Cord Organoid Differentiation Kit and express expected markers via immunostaining. (A) At Day 30, motor neuron markers ISL1 (magenta), HB9 (green), and CHAT (cyan) are expressed, as seen in the upper panel (see insets), as well as motor neuron marker FOXP1 (cyan) and V2a glutamatergic interneuron marker CHX10 (red), as seen in the lower panel (see insets). (B) From Day 75 onwards, the STEMdiff™ spinal cord organoids display astrocyte markers GFAP (magenta) and S100β (green), as well as oligodendrocyte marker MBP (cyan) (see insets). hPSCs = human pluripotent stem cells
Figure 4. STEMdiff™ Spinal Cord Organoids Are Functionally Active
Spinal cord organoids were generated from iPSC line STiPS-M001 and ES cell line H9 using the STEMdiff™ Spinal Cord Organoid Differentiation Kit. At Day 30, the organoids were plated on an MEA (CytoView MEA 48, Axion Biosystems) and activity from 16 electrodes per well were recorded once per week for 5 minutes using a Maestro MEA system (Axion Biosystems). (A) Representative brightfield images of STEMdiff™ spinal cord organoids on the MEA and representative spike heat maps for the corresponding wells (white = 81 and 74 spikes/sec) for STiPS-M001 and H9 are shown respectively. (B) Raster plots of spike activity show the Day 70 STEMdiff™ spinal cord organoids increase in network bursting (pink lines) after 5 weeks of culture on the MEA. (C) Graphs comparing the number of spikes, mean firing rate, synchrony index, and number of active electrodes from Week 1 to 5. The data was normally distributed by D’Agostino & Pearson test and statistics was calculated with paired t-test. ** p ≤ 0.01. iPSC = induced pluripotent stem cell; ES = embryonic stem; MEA = multielectrode array; MFR = mean firing rate
Figure 5. STEMdiff™ Spinal Cord Organoids Express High Levels of Cervical Motor Neuron Markers by qPCR
Spinal cord organoids were generated from hPSCs with the STEMdiff™ Spinal Cord Organoid Differentiation Kit and compared to organoids generated with the STEMdiff™ Dorsal Forebrain Organoid Differentiation Kit. RNA from duplicate organoids was harvested for qPCR analysis. High levels of cervical motor neuron markers are expressed in STEMdiff™ spinal cord organoids (A) at Day 19 and (B) at Day 30, as compared to STEMdiff™ dorsal forebrain organoids. Fold change is reported relative to their parent hPSC and the TBP housekeeping gene control. HOXA4 and HOXA5 graphs use a log10 scale on the y-axis, all other y-axes use a split linear scale. The data is not normally distributed by D’Agostino & Pearson test and statistics were calculated with Mann-Whitney test. **** p ≤ 0.0001. hPSCs = human pluripotent stem cells; qPCR = quantitative polymerase chain reaction
请在《产品说明书》中查找相关支持信息和使用说明,或浏览下方更多实验方案。
本产品专为以下研究领域设计,适用于工作流程中的高亮阶段。探索这些工作流程,了解更多我们为各研究领域提供的其他配套产品。
Thank you for your interest in IntestiCult™ Organoid Growth Medium (Human). Please provide us with your contact information and your local representative will contact you with a customized quote. Where appropriate, they can also assist you with a(n):
Estimated delivery time for your area
Product sample or exclusive offer
In-lab demonstration
配方类别 | Serum-Free |
---|
提升神经元功能的无血清基础培养基
用于从人多能干细胞高效生成背侧前脑神经类器官的细胞培养基试剂盒
用于从hPSCs生成运动神经元的分化试剂盒
扫描二维码或搜索微信号STEMCELLTech,即可关注我们的微信平台,第一时间接收丰富的技术资源和最新的活动信息。
如您有任何问题,欢迎发消息给STEMCELLTech微信公众平台,或与我们通过电话/邮件联系:400 885 9050 INFO.CN@STEMCELL.COM。