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MyoCult™-SF 扩增添加物试剂盒 (人)

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

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产品号 #（选择产品）

产品号 #05980

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

产品优势

支持在无血清条件下对人骨骼肌祖细胞进行衍生和长期扩增。
经培养扩增的骨骼肌祖细胞保持分化潜能。
严格的原材料筛选和质量控制可确保最低的批次间的差异性。
培养扩增的肌源性祖细胞与MyoCult™分化试剂盒兼容。

产品组分包括

MyoCult™-SF 扩增10X添加物（人），2 x 10 mL
MyoCult™-SF 贴附基质，100 µg

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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 A, ACF
Animal component-free collagenase for the digestion of native collagen fibrils
DMEM with 1000 mg/L D-Glucose
Dulbecco's Modified Eagle's Medium (DMEM) with 1000 mg/L D-glucose
Culture Dish, Non-Treated
Sterile, flat-bottom, clear polystyrene non-treated dish with lid; 35, 60, or 100 mm formats
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

在无血清条件下培养原代人骨骼肌祖细胞和人多能干细胞（hPSC）来源的成肌细胞。

MyoCult™-SF 扩增添加物试剂盒 (人) 包含专为成肌细胞衍生与体外扩增优化的组分：MyoCult™-SF 扩增10X添加物（人）和MyoCult™-SF 贴附基质。该添加物需与基础培养基（含1000 mg/L D-葡萄糖的DMEM，产品号 #36253；单独销售）混合以配制MyoCult™-SF扩增培养基。根据所用细胞来源，可选择使用MyoCult™-SF 贴附基质。为方便起见，试剂盒组分也可单独购买。

若使用原代成肌细胞，建议用MyoCult™-SF 贴附基质包被培养器皿。对于hPSC来源的成肌细胞，可使用Corning® Matrigel® hESC-Qualified Matrix。使用MyoCult™-SF扩增培养基培养的成肌细胞与MyoCult™分化试剂盒（人）兼容，可进一步分化为多核肌管结构。

有关使用MyoCult™进行人骨骼肌祖细胞衍生与扩增的详细方案，请参阅产品说明书。

亚型
专用培养基

细胞类型
肌源干/祖细胞

种属
人

应用
细胞培养，扩增

品牌
MyoCult

研究领域
干细胞生物学

制剂类别
无血清

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实验数据

Figure 1. Workflow for the Derivation, Expansion and Differentiation of Human Myogenic Progenitor Cells

Human skeletal muscle tissue is digested into a single cell suspension and plated into MyoCult™-SF Expansion Medium. Myoblasts are then enriched by cell sorting or using a selection kit such as the EasySep™ Human CD56 Positive Selection Kit (Catalog #17855). Purified myoblasts are then culture-expanded or differentiated into myotubes for further studies. Myoblasts can be derived and expanded, using the MyoCult™-SF Expansion Kit (Catalog #05980) and then differentiated using the MyoCult™ Differentiation Kit (Catalog #05965).

Figure 2. Derivation of PAX7+ Myogenic Progenitor Cells in MyoCult™-SF Expansion Medium from Human Skeletal Muscle Tissue

Myogenic progenitor cells were derived from human skeletal muscle tissue and culture-expanded using the MyoCult™-SF Expansion Kit or a serum-containing medium. (A) Following 6 days of expansion, myoblasts were fixed and immunostained for PAX7 (green), a thymine analogue (EdU; red) and nuclei (DAPI; blue). Total percentage of cells expressing (B) PAX7 or (C) PAX7 and EdU were quantified (n = 4). Data was generated and used with permission by Dr. Penny M. Gilbert’s Lab, Department of Biochemistry, University of Toronto.

Figure 3. Long-Term Expansion of Myoblasts were Observed When Using MyoCult™-SF Expansion Kit

Myoblasts were culture-expanded in MyoCult™-SF Expansion Medium or in serum-containing medium. (A) Myoblasts culture-expanded in MyoCult™-SF Expansion Medium displayed superior expansion rate than when cells were expanded in serum-containing medium (n=3). (B) Expansion of myoblasts in MyoCult™-SF Expansion Medium generates a greater yield of CD56+ cells after 9 passages compared to myoblasts cultured in serum-containing medium (P9; n = 2). Data was generated and used with permission by Dr. Penny M. Gilbert’s Lab, Department of Biochemistry, University of Toronto.

Figure 4. Myoblasts Culture-Expanded in MyoCult™-SF Expansion Medium Maintain Differentiation and Transplantation Potential in Mice

Myoblasts derived and expanded in MyoCult™-SF Expansion Medium were further differentiated into myotubes using the MyoCult™ Differentiation Kit (Catalog #05965) at passage 5. (A) Myotubes differentiated from myoblasts were immunostained for myosin heavy chain (MyHC; red) and nuclei (DAPI; blue). (B) Fusion index displaying approximately 60% of total nuclei were found within myotubes expressing MyHC. Each donor is indicated by a yellow square. Data from 5 independent donors.

Figure 5. The MyoCult™-SF Expansion Medium Supports Expansion of iPSC-Derived Myogenic Progenitor Cells

(A) iPSC-derived myogenic progenitor cells displayed long-term expansion when using the MyoCult™ Expansion Kit. iPSC-derived myogenic progenitor cells were generated using Protocol A (Chal et al. from Nature Protocols (2016)) and Protocol B (Xi et al. Cell Rep (2017)). (B) Culture-expanded iPSC-derived myogenic progenitor cells expressed MyHC (red) and MyoD (green).

产品说明书及文档

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。

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

MyoCult™-SF Expansion Supplement Kit (Human)

Catalog #

05980

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

MyoCult™-SF Expansion Supplement Kit (Human)

Catalog #

05980

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

MyoCult™-SF Expansion Supplement Kit (Human)

Catalog #

05980

Lot #

All

Language

English

应用领域

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。

Research Area

Workflow Stages

Workflow Stages for Myogenic Stem and Progenitor Cell Research

Culture and Cryopreservation

Differentiation

Characterization

Workflow Stages for Cardiac and Skeletal Muscle Research

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相关材料与文献

Differentiation of Intrafusal Fibers from Human Induced Pluripotent Stem Cells. A. Col\'on et al. ACS chemical neuroscience 2020

Abstract

Human-based body-on-a-chip" technology provides powerful platforms in developing models for drug evaluation and disease evaluations in phenotypic models. Induced pluripotent stem cells (iPSCs) are ideal cell sources for generating different cell types for these in vitro functional systems and recapitulation of the neuromuscular reflex arc would allow for the study of patient specific neuromuscular diseases. Regarding relevant afferent (intrafusal fibers sensory neurons) and efferent (extrafusal fibers motoneurons) cells in vitro differentiation of intrafusal fiber from human iPSCs has not been established. This work demonstrates a protocol for inducing an enrichment of intrafusal bag fibers from iPSCs using morphological analysis and immunocytochemistry. Phosphorylation of the ErbB2 receptors and S46 staining indicated a 3-fold increase of total intrafusal fibers further confirming the efficiency of the protocol. Integration of induced intrafusal fibers would enable more accurate reflex arc models and application of this protocol on patient iPSCs would allow for patient-specific disease modeling."

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CD4+ T cell activation and associated susceptibility to HIV-1 infection in vitro increased following acute resistance exercise in human subjects. A. K. Holbrook et al. Physiological reports 2019 sep

Abstract

Early studies in exercise immunology suggested acute bouts of exercise had an immunosuppressive effect in human subjects. However, recent data, show acute bouts of combined aerobic and resistance training increase both lymphocyte activation and proliferation. We quantified resistance exercise-induced changes in the activation state of CD4+ T lymphocytes via surface protein expression and using a medically relevant model of infection (HIV-1). Using a randomized cross-over design, 10 untrained subjects completed a control and exercise session. The control session consisted of 30-min seated rest while the exercise session entailed 3 sets × 10 repetitions of back squat, leg press, and leg extensions at 70{\%} 1-RM with 2-min rest between each set. Venous blood samples were obtained pre/post each session. CD4+ T lymphocytes were isolated from whole blood by negative selection. Expression of activation markers (CD69 {\&} CD25) in both nonstimulated and stimulated (costimulation through CD3+ CD28) cells were assessed by flow cytometry. Resistance exercised-induced effects on intracellular activation was further evaluated via in vitro infection with HIV-1. Nonstimulated CD4+ T lymphocytes obtained postexercise exhibited elevated CD25 expression following 24 h in culture. Enhanced HIV-1 replication was observed in cells obtained postexercise. Our results demonstrate that an acute bout of resistance exercise increases the activation state of CD4+ T lymphocytes and results in a greater susceptibility to HIV-1 infection in vitro. These findings offer further evidence that exercise induces activation of T lymphocytes and provides a foundation for the use of medically relevant pathogens as indirect measures of intracellular activation.

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Metformin blunts muscle hypertrophy in response to progressive resistance exercise training in older adults: A randomized, double-blind, placebo-controlled, multicenter trial: The MASTERS trial. R. G. Walton et al. Aging cell 2019 dec

Abstract

Progressive resistance exercise training (PRT) is the most effective known intervention for combating aging skeletal muscle atrophy. However, the hypertrophic response to PRT is variable, and this may be due to muscle inflammation susceptibility. Metformin reduces inflammation, so we hypothesized that metformin would augment the muscle response to PRT in healthy women and men aged 65 and older. In a randomized, double-blind trial, participants received 1,700 mg/day metformin (N = 46) or placebo (N = 48) throughout the study, and all subjects performed 14 weeks of supervised PRT. Although responses to PRT varied, placebo gained more lean body mass (p = .003) and thigh muscle mass (p {\textless} .001) than metformin. CT scan showed that increases in thigh muscle area (p = .005) and density (p = .020) were greater in placebo versus metformin. There was a trend for blunted strength gains in metformin that did not reach statistical significance. Analyses of vastus lateralis muscle biopsies showed that metformin did not affect fiber hypertrophy, or increases in satellite cell or macrophage abundance with PRT. However, placebo had decreased type I fiber percentage while metformin did not (p = .007). Metformin led to an increase in AMPK signaling, and a trend for blunted increases in mTORC1 signaling in response to PRT. These results underscore the benefits of PRT in older adults, but metformin negatively impacts the hypertrophic response to resistance training in healthy older individuals. ClinicalTrials.gov Identifier: NCT02308228.

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