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
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
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
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.
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.
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.
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).
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.
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
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Species | Human |
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Formulation Category | Serum-Free |
人骨骼肌祖细胞(成肌细胞)分化为肌管的培养基
用于人骨骼肌祖细胞(成肌细胞)衍生与扩增的无血清添加物及贴附基质
无动物成分的用于消化天然胶原纤维的胶原酶
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