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AlphaSTEM™ Naïve hPSC Medium

Product Description

Stem Cell Culture Medium | 500ml 

AlphaSTEM™ Naïve hPSC Medium is a serum-free and feeder-free medium formulated for the generation and expansion of naïve pluripotent stem cells.  Utilizing a naturally occurring growth factor, NME7AB , that is expressed in the earliest days of embryogenesis, AlphaSTEM™ Naïve hPSC Medium fully supports the naïve state in human stem cells.  The medium can also be used during reprogramming for high efficient generation of naïve induced pluripotent stem cells.  These cells express pluripotency markers, as well as naïve markers, have two active X chromosomes (female lines) and differentiate down all three germlines.

AlphaSTEM™ Naive hPSC Medium delivers:

  • Enhanced feeder-free reprogramming that is simpler and 20 times more efficient
  • Efficient large-scale expansion using single-cell passaging while maintaining a normal karyotype
  • Superior directed differentiation with improved yield and cell quality

Enhanced Feeder-Free Reprogramming
Despite the desire to move away from the variability and labor associated with MEFs, scientists mistakenly believe they need feeders in order to achieve high efficiency reprogramming. The AlphaSTEM™ System provides high efficiency feeder-free reprogramming with improved cell line survival. Using the Yamanaka factors with AlphaSTEM™ Naive hPSC Medium results in iPSCs at the earliest naïve stem cell state, and the process is simpler and 20 times more efficient.

Efficient Large-Scale Expansion
Cells cultured in AlphaSTEM™ Naive hPSC Medium can be passaged as single cells and used for large-scale expansion in high-capacity flasks resulting in 10-20 fold expansion in four days.  Stem cell expansion is faster, scalable and can be completely automated since it eliminates unwanted spontaneous differentiation, while maintaining a normal karyotype. The media supports hPSC expansion for >70 passages with a normal karyotype.

Superior Directed Differentiation
Naïve stem cells do not have DNA methylation marks that commit the cells to certain developmental decisions. The result is differentiation of cells grown in AlphaSTEM™ Naive hPSC Medium is superior to that of FGF grown cells in terms of quality, quantity and purity with no clonal differentiation bias. 

$295.00

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Data

        B. 

  • A. X- chromosome inactivation in Naïve cells - Cells grown in FGF (left) are showing X chromosome inactivation, where the cells grown in AlphaSTEM™ have no X inactivation, typical of naïve cells.
  • B. Naïve cells have a unique genetic profile - RNA sequencing shows a completely different genetic fingerprint between cells grown in bFGF and cells grown with AlphaSTEM™.


10X- 20X Expansion in 4days

 

User Guide

Keys to Success – Please Read Before Starting

  • AlphaSTEM™ Naive hPSC Medium media does not contain FGF2 and is not compatible with FGF2 media.
  • Culture conditions:  Hypoxic incubator (37°C, 5% CO2, 5% O2).
  • Rho kinase I inhibitor
  • AlphaSTEM™ Naive hPSC Medium is used with AlphaSTEM™ Culture Substrate coated plates.  The medium can be used with other surfaces such as MEFs and Matrigel®; However, growth over these and other surfaces has been shown to compromise naïve characteristics of the resultant cells.
  • The AlphaSTEM™ Culture System uses TrypLE™ dissociated single cells which grow in a uniform monolayer.
  • Passage cells at 75-85% confluence (~3-4 days). Do not overgrow the cells.
  • When first working with naïve cells, it is ideal to have existing naïve stem cells to use as a reference/control.  Minerva offers both male and female naïve stem cells lines.

Download Protocol

 

Publications

A Primitive Growth Factor, NME7AB , Is Sufficient to Induce Stable Naïve State Human Pluripotency; Reprogramming in This Novel Growth Factor Confers Superior Differentiation.

Carter MG, Smagghe BJ, Stewart AK, Rapley JA, Lynch E, Bernier KJ, Keating KW, Hatziioannou VM, Hartman EJ, Bamdad CC. Stem Cells. 2016 Apr;34(4):847-59. doi: 10.1002/stem.2261. Epub 2016 Jan 7.A Primitive Growth Factor, NME7AB , Is Sufficient to Induce Stable Naïve State Human Pluripotency; Reprogramming in This Novel Growth Factor Confers Superior Differentiation.

Abstract
Scientists have generated human stem cells that in some respects mimic mouse naïve cells, but their dependence on the addition of several extrinsic agents, and their propensity to develop abnormal karyotype calls into question their resemblance to a naturally occurring "naïve" state in humans. Here, we report that a recombinant, truncated human NME7, referred to as NME7AB here, induces a stable naïve-like state in human embryonic stem cells and induced pluripotent stem cells without the use of inhibitors, transgenes, leukemia inhibitory factor (LIF), fibroblast growth factor 2 (FGF2), feeder cells, or their conditioned media. Evidence of a naïve state includes reactivation of the second X chromosome in female source cells, increased expression of naïve markers and decreased expression of primed state markers, ability to be clonally expanded and increased differentiation potential. RNA-seq analysis shows vast differences between the parent FGF2 grown, primed state cells, and NME7AB converted cells, but similarities to altered gene expression patterns reported by others generating naïve-like stem cells via the use of biochemical inhibitors. Experiments presented here, in combination with our previous work, suggest a mechanistic model of how human stem cells regulate self-replication: an early naïve state driven by NME7, which cannot itself limit self-replication and a later naïve state regulated by NME1, which limits self-replication when its multimerization state shifts from the active dimer to the inactive hexamer.

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Addtional Minerva publications available HERE

Application Notes & Posters

Download App NoteGeneration of Neural Stem Cells from AlphaSTEM™ Cultured Pluripotent Stem Cells 

Download PosterNovel stem cell growth factor NME7AB dramatically improves iPSC generation, expansion and differentiation 

 

Property Rights, Use Restrictions and Limited License

Property Rights, Use Restrictions and Limited License 

  1. Ownership. Customer acknowledges that the Products provided to Customer embody intellectual property deemed to be of significant value to Minerva Biotechnologies and its licensors, and that such intellectual property may be protected by the law of patents, copyrights, trade secrets, and other laws. Customer acknowledges and agrees that neither this Agreement nor the purchase of the Products by Customer shall be construed as a transfer of any title or the grant of any rights in and to the intellectual property embodied in the Products, owned or licensed by Minerva Biotechnologies. The Products are covered by one or more Minerva patents. Customer has a limited license to use the Products under the patents, subject to the use restrictions and third party licenses in subsections B and C. Customer hereby grants to Minerva Biotechnologies a perpetual, royalty-free, fully paid up, non-exclusive, worldwide, unlimited license to use for any purpose any and all improvements made by Customer to the intellectual property embodied in the Products. Such rights include, without limitation, improvements related to reanimation or thawing, cell functionality, cell differentiation, cell plating, cell improvement assays; or protocols relating to the same. Customer is not granting any rights to and retains all rights to any and all findings and data relating to the performance of the Customer's products, compounds or molecules on the Products.
  2. USE RESTRICTIONS. The Products and/or components of the Products are licensed for internal research purposes only, and may not be used for any other purpose. No other right, express or implied, is conveyed by the sale of the Products. In particular, no right to make, have made, offer to sell, or sell the Products is implied by the sale or purchase of the Products. The license granted herein does not imply or convey the right to use the Products in combination with any other product(s) whose manufacture, sale, or use is covered by any patent. Customer may not transfer the Products to any third party without Minerva Biotechnologies' prior written consent. Customer shall not reverse engineer, modify or otherwise alter the Products in any way. Customer shall not use the Products or any components thereof in humans, in clinical trials or for diagnostic purposes involving human subjects, for any therapeutic use or investigational use, nor for any purpose in contravention of any applicable law, regulation, ordinance, institutional review board approved protocol, or privacy office approval. The Customer cannot sell or otherwise transfer (a) this Product, (b) its components, or (c) materials made by the employment of this Product or its components to a third party or otherwise use this Product or its components or materials made by the employment of this Product or its components for Commercial Purposes.
  3. THIRD-PARTY LICENSES. (i) One or more of the Products may have been generated using intellectual property rights owned by Academia Japan and/or ID Pharma under license agreements with Academia Japan and ID Pharma. The purchase of this Product conveys to the Customer the limited, non-transferable right to use the Product for internal research conducted by the Customer (whether the Customer is an academic or for-profit entity). The Customer cannot sell or otherwise transfer (a) this Product, (b) its components, or (c) materials made by the employment of this Product or its components to a third party or otherwise use this Product or its components or materials made by the employment of this Product or its components for Commercial Purposes. However, the Customer may use differentiated stem cells purchased from Minerva, such as cardiomyocytes, hepatocytes or neural cells, to provide a service. Commercial use of one or more of the Products requires a license from Academia Japan, ID Pharma and/or Minerva Biotechnologies. For license information, please contact Academia Japan at 207 International Science Innovation Building East Wing Kyoto University Yoshida-honmachi, Sakyo-ku, Kyoto, 606-8501 Japan, license@ips-ac.co.jp, ID Pharma at Techno Park Oho, 6 Ohkubo, Tsukuba, Ibaraki, 300-2611 JAPAN, cytotune@dnavec-corp.com and Minerva Biotechnologies at 40 Bear Hill Road, Waltham, MA 02451 license@minervabio.com.
  4. Limited Warranty
    1. Minerva Biotechnologies warrants that its Products conform to the specifications contained in the Certificate of Analysis for the Product shipped to Customer. Minerva Biotechnologies’ sole and exclusive liability under this limited warranty shall be replacement of the defective Products by Minerva Biotechnologies.
    2. Minerva Biotechnologies reserves the right to make changes in design, production, manufacture, or characteristics of the Products or to improve on the Product at any time and in any way, without incurring any obligations to replace or modify any Products previously sold.
    3. Under no circumstances shall Minerva Biotechnologies' liability to Customer exceed the amount paid by Customer for the Products to Minerva Biotechnologies. Minerva Biotechnologies will bear all reasonable shipping costs if the Products are replaced pursuant to this warranty. This warranty does not apply to any defect or nonconformance caused by (i) Customer's use of the Products for a purpose or in a manner other than that for which they were designed or that is permitted or in breach of this Agreement, (ii) the failure by Customer to follow Minerva Biotechnologies' User's Guide for use, storage, and handling of the Products; or (iii) as a result of any other abuse, misuse or neglect of the Products by Customer. This warranty applies only to Customer and not to third parties. This warranty is not assignable.
    4. TO THE FULLEST EXTENT PERMITTED BY APPLICABLE LAW, MINERVA BIOTECHNOLOGIES DISCLAIMS ALL OTHER REPRESENTATIONS, AND WARRANTIES, EXPRESS OR IMPLIED, WITH RESPECT TO THE PRODUCTS, INCLUDING BUT NOT LIMITED TO, ANY IMPLIED WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE OR NON-INFRINGEMENT. CUSTOMER'S SOLE REMEDY FOR BREACH OF WARRANTY IS STATED ABOVE.