STEMCELL Technologies STEMdiff STEMdiff Cardiomyocyte Expansion Kit
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- 新製品
STEMdiff™ Cardiomyocyte Expansion Kit (ST-100-1109) は、ヒト多能性幹細胞（hPSC）由来の初期段階の心筋細胞を安定的に増殖させる無血清培地です。この市場初のキットを使用することで、機能的で高純度のhPSC由来心筋細胞を大量に作製できます。本品は、STEMdiff™ Ventricular Cardiomyocyte Differentiation KitまたはSTEMdiff™ Atrial Cardiomyocyte Differentiation Kitでそれぞれ作製した心室心筋または心房心筋細胞を増殖可能です。
一般的な方法では、大量のhPSC由来心筋細胞を得るために最初に大量のhPSCを増殖し、その後、何回も分化を繰り返します。一方、本品を使用すると、1回の心筋細胞分化で得られる初期段階のhPSC由来心筋細胞を増殖するだけで、簡単に数十億単位の心筋細胞集団に到達できます。このため時間とリソースの節約になります。
本品で増殖した初期段階のhPSC由来心筋細胞は、安定した電気的特性を保持し、継代5回でcTnT陽性率90％を超えます。増殖したhPSC由来心筋細胞は、ハイスループットな薬剤試験、組織工学、再生医療研究に利用できます。
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™ Cardiomyocyte Expansion Kitをもちいて、hPSC由来の初期心筋細胞を血清フリーで増殖できます
- シンプルで操作しやすいワークフローによって心筋細胞の収量をスケールアップできます
- hPSC由来の心房心筋細胞または心室心筋細胞を、最大70倍に増殖させます
- 完全に機能性を備えた心筋細胞を作製し、下流のアッセイにそのまま使用できます
- 純度90%を超える心筋細胞を作製できます
初期心筋細胞の増殖の流れ
Figure 1. Workflow for the Expansion of Early-Stage hPSC-Derived Cardiomyocytes Using the STEMdiff™ Cardiomyocyte Expansion Kit
Expansion of early-stage human pluripotent stem cell (hPSC)-derived cardiomyocytes begins with the differentiation of hPSCs to cardiomyocytes using the STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit or the STEMdiff™ Atrial Cardiomyocyte Differentiation Kit. On Day 11 of differentiation, a beating, confluent monolayer of early-stage cardiomyocytes can be observed. These cardiomyocytes are harvested and replated at low density to initiate expansion using STEMdiff™ Cardiomyocyte Expansion Kit. Expanding hPSC-derived cardiomyocytes can be passaged up to 5 times before switching to STEMdiff™ Cardiomyocyte Maintenance Medium for downstream assays.
データ紹介
hPSC由来心筋細胞を再播種後、1週間でコンフルエンスに到達
Figure 2. Expanding hPSC-Derived Cardiomyocytes Are Confluent One Week After Replating with STEMdiff™ Cardiomyocyte Expansion Kit
Representative images of expanding hPSC-derived ventricular cardiomyocytes after replating at low density (Day 1) using STEMdiff™ Cardiomyocyte Passaging Medium. Cardiomyocyte confluency increases during the 7 day expansion period. On Day 7, a confluent monolayer of beating hPSC-derived cardiomyocytes is observed and the expanded hPSC-derived cardiomyocytes are ready for harvest.
複数のhPSC株由来心筋細胞で増殖を確認
Figure 3. Expansion of hPSC-Derived Cardiomyocytes Across Multiple Cell Lines with STEMdiff™ Cardiomyocyte Expansion Kit
(A) Expansion of hPSC-derived cardiomyocytes was tracked for 5 passages with 4 hPSC lines. Per passage, fold expansion is typically highest in the first 1 – 2 passages. (B) Cumulative fold expansion over 5 passages ranged from 33- to 103-fold expansion and averaged at 72-fold ± 12 (n = 15, 4 hPSC lines).
心筋細胞の純度は、増殖中に上昇する
Figure 4. Cardiomyocyte Purity Increases During Expansion Using STEMdiff™ Cardiomyocyte Expansion Kit
Cardiomyocyte purity was tracked during expansion using STEMdiff™ Cardiomyocyte Expansion Kit for 5 passages on 4 hPSC lines. Throughout expansion, hPSC-derived cardiomyocytes express high levels of cardiac troponin T, and on average show an increase in cardiomyocyte purity during expansion. After 5 passages, cardiomyocyte cultures show an average cTnT purity of 90% ± 2% (n = 15).
増殖後の心筋細胞は、典型的な心臓の電気生理的特性および安定した拍動特性を示す
Figure 5. Post-Expanded hPSC-Derived Cardiomyocytes Exhibit Typical Cardiac Electrophysiology and a Stable Beat Profile
Microelectrode Array (MEA) recordings of post-expanded hPSC-derived cardiomyocytes (using STEMdiff™ Cardiomyocyte Expansion Kit) exhibit a stable beat rate across 4 hPSC lines. The post-expanded hPSC-derived cardiomyocytes were replated onto MEA and maintained in STEMdiff™ Cardiomyocyte Maintenance Medium for 1 week. The average beat rate was 12 ± 1 beats per minute (n = 14, 4 hPSC lines).
