产品号 #07180_C
成分明确的无异源基质，支持人多能干细胞在无血清、无饲养层条件下的生长和分化
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
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CellAdhere™ Dilution BufferDilution buffer for matrix proteins, e.g. Vitronectin XF™
使用玻连蛋白 XF™（一种成分确定且无异源成分的细胞培养基质）支持人多能干细胞的生长和分化。
玻连蛋白 XF™由Nucleus Biologics开发和制造，是康宁Matrigel®的有效替代品。将玻连蛋白 XF™与mTeSR™1, mTeSR™+, TeSR™e8™,或TeSR™-AOF联合使用，为在无饲养层条件下维持人胚胎干细胞（ES）和人诱导多能干细胞（iPS）提供一个明确的培养系统。该系统实现了对培养环境的完全控制，从而获得更稳定的细胞群体，并在后续应用中确保结果的可重复性。
注意:CellAdhere™稀释缓冲液和非组织培养处理的培养器皿（例如产品号#100-0096/27147）需要与玻连蛋白 XF™一起使用，可单独购买。
玻连蛋白 XF™由Nucleus Biologics开发和制造，是康宁Matrigel®的有效替代品。将玻连蛋白 XF™与mTeSR™1, mTeSR™+, TeSR™e8™,或TeSR™-AOF联合使用，为在无饲养层条件下维持人胚胎干细胞（ES）和人诱导多能干细胞（iPS）提供一个明确的培养系统。该系统实现了对培养环境的完全控制，从而获得更稳定的细胞群体，并在后续应用中确保结果的可重复性。
注意:CellAdhere™稀释缓冲液和非组织培养处理的培养器皿（例如产品号#100-0096/27147）需要与玻连蛋白 XF™一起使用，可单独购买。
Data Figures
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
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.
Resources and Publications
Educational Materials (14)
Publications (4)
Non-integrating episomal plasmid-based reprogramming of human amniotic fluid stem cells into induced pluripotent stem cells in chemically defined conditions. Cell cycle (Georgetown, Tex.) 2016 JAN
Abstract
Amniotic fluid stem cells (AFSC) represent an attractive potential cell source for fetal and pediatric cell-based therapies. However, upgrading them to pluripotency confers refractoriness toward senescence, higher proliferation rate and unlimited differentiation potential. AFSC were observed to rapidly and efficiently reacquire pluripotency which together with their easy recovery makes them an attractive cell source for reprogramming. The reprogramming process as well as the resulting iPSC epigenome could potentially benefit from the unspecialized nature of AFSC. iPSC derived from AFSC also have potential in disease modeling, such as Down syndrome or $\$-thalassemia. Previous experiments involving AFSC reprogramming have largely relied on integrative vector transgene delivery and undefined serum-containing, feeder-dependent culture. Here, we describe non-integrative oriP/EBNA-1 episomal plasmid-based reprogramming of AFSC into iPSC and culture in fully chemically defined xeno-free conditions represented by vitronectin coating and E8 medium, a system that we found uniquely suited for this purpose. The derived AF-iPSC lines uniformly expressed a set of pluripotency markers Oct3/4, Nanog, Sox2, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 in a pattern typical for human primed PSC. Additionally, the cells formed teratomas, and were deemed pluripotent by PluriTest, a global expression microarray-based in-silico pluripotency assay. However, we found that the PluriTest scores were borderline, indicating a unique pluripotent signature in the defined condition. In the light of potential future clinical translation of iPSC technology, non-integrating reprogramming and chemically defined culture are more acceptable.
Functionalizing Ascl1 with Novel Intracellular Protein Delivery Technology for Promoting Neuronal Differentiation of Human Induced Pluripotent Stem Cells Stem Cell Reviews and Reports 2016 AUG
Abstract
Pluripotent stem cells can become any cell type found in the body. Accordingly, one of the major challenges when working with pluripotent stem cells is producing a highly homogenous population of differentiated cells, which can then be used for downstream applications such as cell therapies or drug screening. The transcription factor Ascl1 plays a key role in neural development and previous work has shown that Ascl1 overexpression using viral vectors can reprogram fibroblasts directly into neurons. Here we report on how a recombinant version of the Ascl1 protein functionalized with intracellular protein delivery technology (Ascl1-IPTD) can be used to rapidly differentiate human induced pluripotent stem cells (hiPSCs) into neurons. We first evaluated a range of Ascl1-IPTD concentrations to determine the most effective amount for generating neurons from hiPSCs cultured in serum free media. Next, we looked at the frequency of Ascl1-IPTD supplementation in the media on differentiation and found that one time supplementation is sufficient enough to trigger the neural differentiation process. Ascl1-IPTD was efficiently taken up by the hiPSCs and enabled rapid differentiation into TUJ1-positive and NeuN-positive populations with neuronal morphology after 8 days. After 12 days of culture, hiPSC-derived neurons produced by Ascl1-IPTD treatment exhibited greater neurite length and higher numbers of branch points compared to neurons derived using a standard neural progenitor differentiation protocol. This work validates Ascl1-IPTD as a powerful tool for engineering neural tissue from pluripotent stem cells.
Chemically defined conditions for human iPSC derivation and culture. Nature methods 2011 MAY
Abstract
We re-examine the individual components for human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) culture and formulate a cell culture system in which all protein reagents for liquid media, attachment surfaces and splitting are chemically defined. A major improvement is the lack of a serum albumin component, as variations in either animal- or human-sourced albumin batches have previously plagued human ESC and iPSC culture with inconsistencies. Using this new medium (E8) and vitronectin-coated surfaces, we demonstrate improved derivation efficiencies of vector-free human iPSCs with an episomal approach. This simplified E8 medium should facilitate both the research use and clinical applications of human ESCs and iPSCs and their derivatives, and should be applicable to other reprogramming methods.
更多信息
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Legal Statement: Vitronectin XF is developed and manufactured by Nucleus Biologics, and Vitronectin XF™ is a trademark of Nucleus Biologics. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
