STEMdiff™ 肌源祖细胞补充试剂盒

用于将人多能干细胞（hPSCs）分化为肌源祖细胞的无血清补充剂

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产品号 #（选择产品）

产品号 #100-0151_C

用于将人多能干细胞（hPSCs）分化为肌源祖细胞的无血清补充剂

产品优势

可在多个人多能干细胞系中实现高效分化为肌源祖细胞
PSC衍生的肌管显示出有序的肌节蛋白结构和收缩功能
优化的试剂可支持整个从人PSC分化为肌源祖细胞的工作流程

产品组分包括

STEMdiff™ 肌源祖细胞补充剂 A，240 μL
STEMdiff™ 肌源祖细胞补充剂 B，240 μL
STEMdiff™ 肌源祖细胞补充剂 C，240 μL
STEMdiff™ 肌源祖细胞补充剂 D，60 mL

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要查看实验方案所需的所有配套产品，请参阅《实验方案与技术文档》。

Collagenase Type IV (1 mg/mL)
Cell dissociation reagent
D-PBS (Without Ca++ and Mg++)
Dulbecco’s phosphate-buffered saline without calcium and magnesium
Y-27632 (Dihydrochloride)
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
mTeSR™1
cGMP, feeder-free maintenance medium for human ES and iPS cells
Labeling Antibodies
Compatible antibodies for purity assessment of isolated cells

总览

实验数据

产品说明书及文档

应用领域

总览

STEMdiff™ 肌源祖细胞补充试剂盒由一组无血清补充剂组成，用于将人胚胎干（ES）细胞和诱导多能干（iPS）细胞分化为肌源祖细胞。这些细胞可通过肌源性细胞标志物（如 CD56 和 CD82）进行鉴定，并可使用 MyoCult™-SF 扩增补充试剂盒（人；目录号 #05980）进行连续传代培养超过 5 代，随后使用 MyoCult™ 分化试剂盒（人；目录号 #05965）进一步高效分化为具有功能性的多核 MyHC⁺ 肌管。这些肌管可用于多种后续应用与分析。

该试剂盒兼容使用以下培养系统维持的人 ES 和 iPS 细胞：mTeSR™ 1（目录号 #85850）、mTeSR™ Plus（目录号 #100-0276）或 TeSR™-E8™（目录号 #05990）

亚型
添加剂

细胞类型
肌源干/祖细胞

种属
人

应用
细胞培养，分化

品牌
STEMdiff

研究领域
疾病建模，干细胞生物学

制剂类别
无血清

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实验数据

Cell images represinting different stages of myogenic differentiation use STEMdiff Myogenic Progenirot Supplement kit.

Figure 1. Schematic of Myogenic Induction Using the STEMdiff™ Myogenic Progenitor Supplement Kit

Prior to differentiation (Day -1), human pluripotent stem cells (hPSCs) are plated in Corning® Matrigel®-coated plates. On Day 0, differentiation is initiated and continued by performing daily media changes with STEMdiff™ Myogenic Progenitor Supplement A at Days 0 - 2, STEMdiff™ Myogenic Progenitor Supplement B at Days 2 - 4, STEMdiff™ Myogenic Progenitor Supplement C at Days 4 - 6, and STEMdiff™ Myogenic Progenitor Supplement D for the remainder of the protocol (Days 6 - 30). Upon completion of the 30-day protocol, the culture contains an abundance of PAX7⁺MyoD⁺ myogenic progenitors that can be further expanded using MyoCult™-SF Expansion Supplement Kit (Human; Catalog #05980) for use in downstream applications.

Heatmap of differential gene expression for myogenic progenitors differentiated from embryonic stage.

Figure 2. hPSCs Differentiated Using STEMdiff™ Myogenic Progenitor Kit Demonstrate Transcript Patterns of Embryonic Muscle Development

hPSCs (H9 cell line) were differentiated using the STEMdiff™ Myogenic Progenitor Kit and harvested at indicated time points for transcript analysis via qPCR; transcription was normalized against 18s as the housekeeping gene. The resulting heatmap indicates transcriptional changes to genes that mark specific stages of muscle development (mesoderm, paraxial mesoderm, somites, and myogenesis) and demonstrates a cellular trajectory that resembles embryonic muscle development.

Flow cytometry analysis of myogenic progenitors generated using the STEMdiff™ Myogenic Progenitor Kit.

Figure 3. STEMdiff™ Myogenic Progenitor Kit Generates Expandable hPSC-Derived Myogenic Progenitors

(A) Representative image of proliferating sub-cultured hPSC-derived myogenic progenitors generated using the STEMdiff™ Myogenic Progenitor Kit. (B) hPSC-derived myogenic progenitors harvested at passage 1 and 5 expressed human myoblast markers CD56 and CD82. (C) Expansion rates of hPSC-derived myogenic progenitors (hSPC-MP) over 5 passages across multiple hPSC lines are comparable to human primary myoblasts. Error bars represent standard error of mean, n = 3.

Microscopy images of different hPSC cell lines differentiated to myogenic progenitors using STEMdiff™ Myogenic Progenitor Kit and stained for myogenic markers MgHC and MYOD.

Figure 4. STEMdiff™ Myogenic Progenitor Kit Facilitates Efficient Differentiation of hPSC-Derived Myogenic Progenitors into hPSC-Derived Myotubes

(A) hPSC-derived myogenic progenitors were generated from multiple hPSC lines using the STEMdiff™ Myogenic Progenitor Kit and then induced to differentiate into myotubes using MyoCult™ Differentiation Medium (Human; Catalog #05965). After 8 days, myotubes were fixed and stained for MyHC and MyoD. (B) Multiple hPSC cell lines differentiated and induced using said method exhibited high fusion indices similar to primary human myoblasts (Numbers in bars represent the n number and dots represent technical replicates).

Microscopy image and electrophysiology of myotubes differentiated from hPSC-generated myogenic progenitors using STEMdiff™ Myogenic Progenitor Kit.

Figure 5. hPSC-Derived Myotubes Generated Using STEMdiff™ Myogenic Progenitor Kit Are Functionally Contractile

hPSC-derived myogenic progenitors were generated using the STEMdiff™ Myogenic Progenitor Kit and then induced to differentiate into myotubes using MyoCult™ Differentiation Medium. (A) hPSC-derived myotubes were stained for alpha-actinin and displayed organized sarcomeric structures as indicated by the zoomed-in area. (B) Spontaneous field potential recordings of hPSC-derived myotubes using microelectrode assay plate indicated that the derived myotubes are contractile. (C) Representative video of contracting hPSC-derived myotubes.