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NutriFreez® D10 Cryopreservation Medium Zoom

NutriFreez® D10 Cryopreservation Medium

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A chemically defined, animal component-free, protein-free cryopreservation solution

High recovery post thaw

cGMP-manufactured

Validated with various cell types and applications

Drug Master File available

Name SKU Size
NutriFreez® D10 Cryopreservation Medium 05-713-1A 500 mL
NutriFreez® D10 Cryopreservation Medium 05-713-1B 100 mL
NutriFreez® D10 Cryopreservation Medium 05-713-1E 50 mL
NutriFreez® D10 Cryopreservation Medium 05-713-1C 20 mL
NutriFreez® D10 Cryopreservation Medium 05-713-1D 10 mL

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Description

Details

Product Overview

NutriFreez® D10 Cryopreservation Medium is a ready-to-use, animal component-free, serum-free, and protein-free cell freezing solution optimized for multiple cell lines.  When culturing cells in a serum-free environment, it is essential to also maintain serum-free conditions during cryopreservation.  

NutriFreez® D10 Cryopreservation Medium has been demonstrated to result in high rates of cell viability, proliferation, adherence (in relevant lines), and bioactivity/ expression following freezing and thawing.  Superior results were obtained both in comparison with serum-containing freezing media as well as competing serum-free products, making this an ideal product for both serum-containing and serum-free applications.

NutriFreez® D10 Cryopreservation Medium has been extensively tested on multiple cell types, including:

  • Human Mesenchymal Stem Cells (hMSC), from various sources (e.g. bone marrow (BM-MSC), adipose tissue (AT-MSC), umbilical cord tissue (UC-MSC) derived and dental pulp tissue (hMSC –DP)
  • Human Embryonic Stem Cells (hESC), Induced Pluripotent Stem Cells (iPSC)
  • Human Peripheral Blood Mononuclear Cells (PBMC’s)
  • Human Endothelial Cells (EC)
  • T cells, including Chimeric Antigen Receptors (CART) cells and Tumor-Infiltrating Lymphocytes (TIL’s)
  • Neuron Cells
  • Hybridomas
  • CHO cells
  • Vero Cells
  • Multiple mammalian cell lines: MRC-5, HEK-293, HepG2, HeLaBSC-1, BGM3T3, MA-10BHK-21 as well as other extremely sensitive cell types

 

NutriFreez® D10 Cryopreservation Medium Features

  • Cryoprotective formulation designed to minimize dehydration effects
  • Effective maintenance of cell viability, adhesion, and bioactivity
  • Ready-to-use solution, simple protocol
  • Suitable for use in cell banks and biopharmaceutical applications
  • Animal component-free, serum-free, protein-free solution
  • cGMP Manufactured
  • Drug Master File available

Results of a cryopreservation comparison performed at the Research and Development Center for Cell Therapy (Japan) on human pluripotent stem cells (hESC and hiPSC) show that NutriFreez® D10 Cryopreservation Medium* presents best recovery rate for hPSCs after thawing (Nishishita et al. Am J Stem Cells. 2015).

Furthermore, an evaluation of different cryopreservation agents for mesenchymal stem cells (MSC) performed at Ottawa Hospital Research Institute (Canda) shows that MSC cryopreserved in NutriFreez® D10 Cryopreservation Medium exhibit the best post-thaw viability (Salkhordeh et. al. Cytotherapy, May 2018).

* NutriFreez® brand replaces the brand CryoStem™

H1 hESC BGO1V/hOG cels (Variant hESC hOct4-GFP reporter cells)

1 hour post-thawing

Day 1 Post-thawing

Day 4 Post-thawing

hESC were frozen in NutriFreez® D10 Cryopreservation Medium and were thawed into NutriStem® hPSC XF on Matrigel. The cells maintain excellent attachment ability as well as growth performance.

Specifications

Specifications

Storage Conditions 2-8°C
Shipping Conditions Cold Pack
Instructions for Use

Cryopreservation of serum-free cell cultures 

  1. For freezing adherent cells, detach cells using the appropriate dissociation solution. (For freezing of cells in suspension, skip to step 2.)
  2. Centrifuge to pellet the cells at 200 - 300 x g for 3 to 5 minutes.
  3. Suspend the cell pellet in cold NutriFreez® D10 Cryopreservation Medium at a concentration of 1 to 10 million cells/mL.
  4. Freeze the cells gradually (1 to 2°C per minute) and store them in liquid nitrogen.
  5. Viability and recovery of cryopreserved cells should be evaluated 24 hours after storage of vials in liquid nitrogen by following the thawing procedure outlined below.

 

Thawing cryopreserved cells 

  1. Thawing should be performed at 37°C.
  2. Immediately after thawing, suspend the cells in serum-free growth medium at a ratio of at least 1:10.
  3. Centrifuge and suspend in growth medium as desired.
  4. Culture the cells according to the recommended seeding density.

References

references

Pluripotent Stem Cells
  1. L. E. Wiersma, et al. Large-Scale Engineering and Cryopreservation of hiPSC-Derived Nephron Sheets. Stem Cell Research & Therapy,  2021. DOI: https://doi.org/10.21203/rs.3.rs-1116352/v1
  2. M. Breunig, et al. Differentiation of human pluripotent stem cells into pancreatic duct-like organoids. STAR Protocols, 2021, https://doi.org/10.1016/j.xpro.2021.100913.
  3. S.E. Strawbridge, et al. Deriving Human Naïve Embryonic Stem Cell Lines from Donated Supernumerary Embryos Using Physical Distancing and Signal Inhibition. Methods in Molecular Biology, Humana, https://doi.org/10.1007/978-1-0716-1908-7_1
  4. E. Balbasi , et al. Mouse Embryonic Stem Cell Culture in Serum-Containing or 2i Conditions. Methods in Molecular Biology. Springer, 2021. https://doi.org/10.1007/7651_2021_438
  5. D. Khan, et al. Testing the Stability of Drug Resistance on Cryopreserved, Gene-Engineered Human Induced Pluripotent Stem Cells. Pharmaceuticals 2021, https://doi.org/10.3390/ph14090919
  6. B. Liu, et al. Chemically defined and xeno-free culture condition for human extended pluripotent stem cells. Nat Commun (2021). https://doi.org/10.1038/s41467-021-23320-8
  7. R.R. Annand, Generation of Human iPSCs by Reprogramming with the Unmodified Synthetic mRNA. In: Hu K. (eds) Nuclear Reprogramming. Methods in Molecular Biology, (2021) vol 2239. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1084-8_11
  8. J. Selberg, et al. Machine Learning-Driven Bioelectronics for Closed-Loop Control of Cells. Adv. Intell. Syst. 2020, DOI: 10.1002/aisy.202000140
  9. Z. Danhong et al. Method of Cryopreservation of Stem Cell-Derived Retinal Pigment Epithelial Cells on Polymeric Substrate. Application Number: 16/658457, Publication Date: 02/13/2020
  10. M. G. Hernandez, et al.  AAV-Mediated Gene Delivery to 3D Retinal Organoids Derived from Human Induced Pluripotent Stem Cells. Int. J. Mol. Sci. 2020, 21, 994; doi:10.3390/ijms21030994
  11. S. Kumar, et al. Highly efficient induced pluripotent stem cell reprogramming of cryopreserved lymphoblastoid cell lines. Journal of Biological Methods | 2020 | Vol. 7(1) | e124 DOI: 10.14440/jbm.2020.296
  12. Kuznitsov-Yanovsky L., Mayshar Y., Ben-Yosef D. (2019) Modeling FXS: Human Pluripotent Stem Cells and In Vitro Neural Differentiation. In: Ben-Yosef D., Mayshar Y. (eds) Fragile-X Syndrome. Methods in Molecular Biology, vol 1942. Humana Press, New York, NY
  13. G. Gagliardi et at. Characterization and Transplantation of CD73-Positive Photoreceptors Isolated from Human iPSC-Derived Retinal Organoids. Stem Cell Reports j Vol. 11 j 665–680 j September 11, 2018
  14. M. Amit, J. Itskovitz-Eldor Single cells pluripotent stem cells in a suspension culture. US Patent App. 16/055,110, 2018
  15. G. Guo, et al. Epigenetic resetting of human pluripotency." bioRxiv (2017): 146712.
  16. S. Reichman et al. Generation of Storable Retinal Organoids and Retinal Pigmented Epithelium from Adherent Human iPS Cells in Xeno‐Free and Feeder‐Free Conditions.STEM CELLS 35.5 (2017): 1176-1188.
  17. F. Pistollato, et al. Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing. JoVE (Journal of Visualized Experiments) 124 (2017): e55702-e55702.
  18. L. Tian et al. Efficient and controlled generation of 2D and 3D bile duct tissue from human pluripotent stem cell-derived spheroids." Stem Cell Reviews and Reports 12.4 (2016): 500-508.
  19. J. Chal et al. Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro.Nature protocols 11.10 (2016): 1833-1850.
  20. M. S. Y Hiew. Generation and Microrna Expression Profiling of Colorectal Cancer Cell-Derived Induced Pluripotent Cancer Cells. Diss. UTAR, 2016.
  21. N. Nishishita et al. An effective freezing/thawing method for human pluripotent stem cells cultured in chemically-defined and feeder-free conditions. American journal of stem cells 4.1 (2015): 38.
  22. Z. Danhong, et al. Method of cryopreservation of stem cell-derived retinal pigment epithelial cells on polymeric substrate.U.S. Patent Application No. 14/114,182, 2014
  23. C. Martin et al. Highly Efficient microRNA-Enhanced mRNA Reprogramming of Diseased Human Fibroblasts in a Feeder-Free Culture System. STEMGENT Application Note, 2014
  24. C. Buensuceso et al. Induced pluripotent stem cells prepared from human kidney-derived cells . US Patent 20140073049 A1, 2014
  25. R. Sivapatham, X.Zeng. Generation and Characterization of Patient-Specific Induced Pluripotent Stem Cell for Disease Modeling. Methods in Molecular Biology, 2014
  26. L. Tian, N. Prasad, Y. Jang. In Vitro Modeling of Alcohol-Induced Liver Injury Using Human-Induced Pluripotent Stem Cells. Methods in Molecular Biology, 2014
  27. C. Buensuceso et al. Induced pluripotent stem cells from human umbilical cord tissue-derived cells. US Patent 20130157365, 2013
  28. S. Hikita et al. Methods of Culturing Retinal Pigmented Epithelium Cells, Including Xeno-Free Production, RPE Enrichment, and Cryopreservation. US Patent 20130196369 A1, 2013
  29. N. Nishishita et al. Generation of Virus-Free Induced Pluripotent Stem Cell Clones on a Synthetic Matrix via a Single Cell Subcloning in the Naïve State. PLoS ONE 7(6): e38389. doi:10.1371/journal.pone.0038389 , 2012
  30. F. Pistollato et al. Standardization of pluripotent stem cell cultures for toxicity testing. Vol. 8, No. 2 , Pages 239-257 (doi:10.1517/17425255.2012.639763), 2012
Mesenchymal Stem Cells
  1. N. Aydoğdu, et al. Isolation, Culture, Cryopreservation, and Preparation of Skin-Derived Fibroblasts as a Final Cellular Product Under Good Manufacturing Practice–Compliant Conditions. Methods in Molecular Biology. (2020) Springer, New York, NY. https://doi.org/10.1007/7651_2020_333
  2. N. Aydoğdu, et al. Isolation, Culture, Cryopreservation, and Preparation of Umbilical Cord-Derived Mesenchymal Stem Cells as a Final Cellular Product Under Good Manufacturing Practices–Compliant Conditions. Methods in Molecular Biology. (2020) Springer, New York, NY. https://doi.org/10.1007/7651_2020_332
  3. M. Paz Quesada et al., Safety and Biodistribution of Human Bone Marrow-Derived Mesenchymal Stromal Cells Injected Intrathecally in Non-Obese Diabetic Severe Combined Immunodeficiency Mice: Preclinical Study Tissue Eng Regen Med, 03 July 2019, https://doi.org/10.1007/s13770-019-00202-1
  4. A.M Lyness et al. Evaluation of a Vial Adaptor to Ensure Safe and Efficient Needle-Free Transfer of Cells Post-Cryopreservation in Preparation for Drug DeliveryCytotherapy, Volume 21, Issue 5, Supplement, May 2019, Page S38
  5. M. Salkhordeh et al. Evaluation of Different Cryopreservation Agents for Mesenchymal Stem Cell as Final Study Product. Cytotherapy, Volume 20, Issue 5, Supplement, May 2018, Page S50
  6. Kundrotas, Gabrielis, et al. Identity, proliferation capacity, genomic stability and novel senescence markers of mesenchymal stem cells isolated from low volume of human bone marrow. Oncotarget 7.10 (2016): 10788.
  7. Kundrotas, Gabrielis. Investigation of human mesenchymal stem cell genetic characteristics to ensure therapy safety. Diss. Vilnius University, 2016.

Immune cells
  1. F Wang et al. Allogeneic Expanded Human Peripheral NK Cells Control Prostate Cancer Growth in a Preclinical Mouse Model of Castration-Resistant Prostate Cancer. Journal of Immunology Research, 2022
  2. L. Xiao, et al. Spectral Markers for T Cell Death and Apoptosis—A Pilot Study on Cell Therapy Drug Product Characterization Using Raman Spectroscopy. Journal of Pharmaceutical Sciences, 2021, https://doi.org/10.1016/j.xphs.2021.08.005.
  3. R. E. Burnham, et al. Human serum albumin and chromatin condensation rescue ex vivo expanded γδ T cells from the effects of cryopreservation. Cryobiology, 2021, ISSN 0011-2240, https://doi.org/10.1016/j.cryobiol.2021.01.011.
  4. N. Milevoj et. al. A combination of electrochemotherapy, gene electrotransfer of plasmid encoding canine IL-12 and cytoreductive surgery in the treatment of canine oral malignant melanoma. Research in Veterinary Science, volume 122, February 2019, Pages 40-49

Other Cell Lines
  1. Y. Konno. et al. A potential preventive method for scar stenosis after esophageal endoscopic mucosal resection using human amniotic epithelial cells in a porcine model. DEN Open, Volume2, Issue1, 2022
  2. D. Pamies, et al. Guidance document on Good Cell and Tissue Culture Practice 2.0 (GCCP 2.0). ALTEX. 2021 Dec 9. doi: 10.14573/altex.2111011.
  3. Y. Jing-Yan, et al. Retinal Protection by Sustained Nanoparticle Delivery of Oncostatin M and Ciliary Neurotrophic Factor Into Rodent Models of Retinal Degeneration. Translational Vision Science & Technology August 2021, Vol.10, 6. doi:https://doi.org/10.1167/tvst.10.9.6
  4. L.Klipfel, et al. A Splice Variant inSLC16A8Gene Leads to LactateTransport Deficit in Human iPSCell-Derived Retinal Pigment Epithelial Cells. Cells 2021,10,  179.https://doi.org/10.3390/cells10010179
  5. S. Reichman, et al. Compositions and Methods for Efficient Amplification of Retinal Progenitors Cells. US Patent App. 16/976581, 01/07/2021
  6. X.Jiang, et al. Mitochondria Dynamically Transplant into Cells in Vitro and in Mice and Rescue Aerobic Respiration of Mitochondrial DNA-Depleted Motor Neuron NSC-34. Journal of Biomedical Science and Engineering, 13, 203-221 (2020). doi: 10.4236/jbise.2020.139019.
  7. A. C. Brown et al. Long-Term Culture of Nephron Progenitor Cells Ex Vivo. In: Vainio S. (eds) Kidney Organogenesis. Methods in Molecular Biology, vol 1926. Humana Press, New York, NY
  8. E. Nigro, E. Soprana, A. Brini, et l., An Antitumor Cellular Vaccine Based on a Mini-Membrane IgE. J Immunol 188:103-110 (2012).
  9. G. Di Lullo, E. Soprana, M. Panigada, et al. The combination of marker gene swapping and fluorescence-activated cell sorting improves the efficiency of recombinant modified vaccinia virus Ankara vaccine production for human use. J. of Virological Methods, 163, Issue 2: 195–204 (2010)
  10. Amit, Michal, and Joseph Itskovitz-Eldor. Novel methods and culture media for culturing pluripotent stem cells. U.S. Patent Application 13/821,244.
  11. De Falco, Elena, et al. A standardized laboratory and surgical method for in vitro culture isolation and expansion of primary human Tenon’s fibroblasts.Cell and tissue banking 14.2 (2013): 277-287.

* The CryoStem™ brand is replaced by the brand NutriFreez®

Documentation

Materials Safety Data Sheet

Manuals and Protocols

Product Literature

Technical Reports

Frequently Asked Questions

*The CryoStem™ brand is replaced by the brand NutriFreez®

Certificate of Analysis

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COA's can be downloaded from Sartorius's Certificates Portal.

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Reviews

Customer Reviews (6)

Very nice productReview by Raquel Brandao Haga
Quality
I have used this medium to freeze mouse fibroblasts and it has worked very well. It was very easy to use and my cells looked very health and happy after I thawed them. (Posted on 3/13/2020)
Very reliable for organoids freezeReview by Shengzhen Guo
Quality
Mouse tumoroids were dissociated from the 3D culture embedded in matrigel. Two freezing medium were compared for freezing down and recovery. One is from Gibco (12648010), which I normally used before, and the second one is NutriFreez. Both performed very nice. Since the organoids were cultured without any serum so would prefer to use Nutrifreez in the future. (Posted on 3/6/2020)
Great product for stem cell long term freezing!Review by Bar Philosof
Quality
I use it to freeze cancer stem-like cell, and it is a great product. (Posted on 8/7/2019)
great productReview by Amira gepstein
Quality
I freeze IPS cells with great succses. Its easier and more effective then freezing with

FBS(20%+ DMSO(10%or 20%). (Posted on 8/7/2019)
This is a recommendable product.Review by Zhiyuan Lv
Quality
This is my first use, and it will be my only choice. It is convenient to use and increases of cell viabilities. (Posted on 8/7/2019)
-Review by Shaked Eliyahu
Quality
I use this product regularly with cells which are not supposed to receive serum. It is very good and the cells recover quickly. I really recommend this product. (Posted on 8/7/2019)

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