STEMCELL Technologies STEMdiff STEMdiff Myogenic Progenitor Supplement Kit
- 研究用
STEMdiff™ Myogenic Progenitor Supplement Kit（ST-100-0151）は、ヒト胚性幹（ES）細胞および人工多能性幹（iPS）細胞を筋原性前駆細胞に分化させるための、無血清サプリメントです。
得られる筋原性前駆細胞は、CD56やCD82などの筋形成細胞マーカーで特徴づけられ、MyoCult™-SF Expansion Supplement Kit（ST-05980）をもちいて5継代以上の拡大培養をおこなうことができます。さらに、MyoCult™ Differentiation Kit（ST-05965）をもちいて機能性の多核MyHC+筋管（myotube）に高効率で分化させることができます。これらの筋管は、さまざまな下流のアプリケーションや分析に使用できます。
本品は、mTeSR™1（ST-85850）、mTeSR™ Plus（ST-100-0276）、またはTeSR™-E8™（ST-05990）のいずれかで維持されているヒトES/iPS細胞からの分化に適しています。
2018/05/14 12:00の製品情報
本製品は研究目的にのみ使用し、人や動物の医療用・臨床診断用・食品用としては使用しないようにご注意ください。
製品の特長
STEMdiff™ Myogenic Progenitor Supplement Kitをもちいて、ヒトES/iPS細胞から効率的に筋原性前駆細胞を誘導できます
- ヒト多能性幹細胞（hPSC）から、細胞株に依存せずに筋原性前駆細胞へ効率的に分化できます
- 組織化されたサルコメアタンパク質と収縮性を示す筋管が得られます
- hPSC由来筋原性前駆細胞ワークフローの全体をサポートするよう最適化されています
筋誘導の流れ
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.
データ紹介
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).
