Nutristem hPSC XF Medium

Nutristem hPSC XF Medium

Defined, xeno-free, serum-free medium
Designed for optimal growth and expansion of human iPS and hES cells

nameSKUsize
NutriStem® hPSC XF Medium, 500mL05-100-1A500 mL
NutriStem® hPSC XF Medium, 100mL05-100-1B100 mL
NutriStem® hPSC XF Medium (Growth Factor-Free)06-5100-01-1A500 mL
  • Description
  • Specifications
  • References
  • Documentation
  • Reviews (0)

Description

NutriStem hPSC Overview

Figure: 1 Human Embryonic Stem Cells (H1, passage 6) were seeded in 96-well plates (Matrigel coated) in BI’s NutriStem hPSC XF and competitor’s media. Stem cell media were changed every 24 hours. Number of cells was determined using CyQuant™ cell proliferation assay kit.

NutriStem® hPSC XF Medium is a defined, xeno-free, serum-free cell culture medium designed to support the growth and expansion of human induced pluripotent stem (hiPS) and human embryonic stem (hES) cells. Nutristem hPSC is widely published and numerous protocols have been established for applications that incorporate the complete cell culture process from derivation to differentiation. NutriStem hPSC XF Medium allows for pluripotent stem cells to be cultured in a completely xeno-free system, without the need for high levels of basic FGF and other stimulatory growth factors and cytokines. The superior cell attachment and proliferation NutriStem hPSC XF Medium provides makes it an excellent medium choice to permit high-throughput screening applications. The reliable and consistent cellular morphology and growth of cells cultures in NutriStem hPSC XF Medium increases experimental reproducibility and has been shown in long-term growth of over 50 passages whilst preserving normal karyotypes.

NutriStem hPSC XF Features

  • Defined, serum-free, and xeno-free
  • Allows for flexibility to derived cells with a variety matrices
  • Allows for weekend-free culture
  • FDA Drug Master File (DMF) available, produced under cGMP
  • Enables efficient expansion and growth of hES and hiPS cells in feeder-free culture systems
  • Low-protein formulation that contains stable L-alanyl-L-glutamine and HSA
  • Extensively tested and widely used on multiple hES and iPS cell lines, including H1, H9.2, I6, I3.2, and CL1

Sample Data

Cell morphology

Figure: Normal Colony Morphology. H1 hES cells (top panel) and ACS-1014 hiPS cells (bottom panel) cultured in NutriStem® hPSC XF Medium on Matrigel-coated plates display colony morphologies typical of normal feeder-free hES and hiPS cell cultures, including a uniform colony of tightly compacted cells and distinct colony edges.

 

 

 

 

 

 

 

 

 

 

 

 

 

Immunostaining

Figure: H1 cell morphology and immunofluorescence analysis of hESC markers red SSEA-4, green OCT4 and blue DAPI. H1 cells stained positive for the expression of pluripotency markers.

 

 

 

 

 

 

 

 

 

 

 

 

Embryoid body formation

Figure: Embryoid bodies (EBs) were generated from H9.2 hES cells cultured for 16 passages in NutriStem® hPSC XF Medium on Matrigel matrix as an evaluation of pluripotency. The pluripotent H9.2 cells were suspended in serum-supplemented medium, where they spontaneously formed EBs containing cells of embryonic germ layers. The following cell types were identified by examination of the histological sections of 14-day-old EBs stained with H&E: (A) neural rosette (ectoderm), (B) neural rosette stained with Tubulin, (C) primitive blood vessels (mesoderm), and (D) megakaryocytes (mesoderm).

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Figure: H9.2 hES cells were cultured for 11 passages in NutriStem® hPSC XF Medium using a human foreskin fibroblast (HFF) feeder layer. The hES cells were subsequently injected into the hind leg muscle of SCID-beige mice for in vitro evaluation of pluripotency. The following tissues from all three germ layers were identified in H&E-stained histological sections of the teratoma 12 weeks post-injection: (A) cartilage (mesoderm), (B) epithelium (endoderm), and (C) neural rosette (ectoderm).

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Instructions for Use  

  • Upon thawing, the medium may be stored at 2 – 8ºC for 10 to 14 days
  • Media should be aliquotted into smaller working volumes to avoid repeated freeze/thaw cycles
  • Avoid exposure to light

Note: A common feeder-free basement membrane matrix is Matrigel, which is not xeno-free. Effective xeno-free alternatives to Matrigel is recombinant laminin, such as LaminStem(R) 521 (BI Cat. No. 05-753-1F) which has been validated to successfully culture human ES and iPS cells using NutriStem® hPSC XF medium.

Legal : NutriStem® hPSC XF is registered as an In-vitro diagnostic (IVD) medical device. NutriStem® is a registered trademark of Biological Industries​.

A Drug Master File (DMF) for NutriStem® hPSC XF is available.

Additional information

Brand

Form

Liquid

Storage Conditions

Store at -20ºC

Shipping Conditions

Dry Ice

Quality Control

NutriStem® hPSC is routinely tested for optimal maintenance and expansion of undifferentiated hESCs. Additional standard evaluations are pH, osmolality, endotoxins and sterility tests.

Growing Methods of hESC and iPSC (Derivation, Expansion, Scaling up, and Suspensions)

  1. O.M. Russell et al. Preferential amplification of a human mitochondrial DNA deletion in vitro and in vivo. Scientific Reports, volume 8, Article number: 1799 (2018)
  2. J. Rosati et al. Production and characterization of human induced pluripotent stem cells (iPSCs) from Joubert Syndrome: CSSi001-A (2850). Stem Cell Research, Volume 27, March 2018, Pages 74–77
  3. Maroof M Adil, David V Schaffer. Expansion of human pluripotent stem cells. Current Opinion in Chemical Engineering 2017, 15:24–35
  4. Tateno, H. et al. Development of a practical sandwich assay to detect human pluripotent stem cells using cell culture media Regenerative Therapy, Volume 6, June 2017, Pages 1–8
  5. Baker, D. et al. Detecting Genetic Mosaicism in Cultures of Human Pluripotent Stem Cells Stem Cell Reports, 2016
  6. Vega-Crespo, A., et al. Investigating the functionality of an OCT4-short response element in human induced pluripotent stem cells. Molecular Therapy — Methods & Clinical Development 3, Article number: 16050 (2016)
  7. Y.Y. Lipsitz, P.W. Zandstra, Human pluripotent stem cell process parameter optimization in a small scale suspensionbioreactor. BMC Proceedings, 9(Suppl 9), O10, 2015
  8. S. Gregory et al. Autophagic response to cell culture stress in pluripotent stem cells. Biochemical and Biophysical Research Communications, doi:10.1016/j.bbrc.2015.09.080, 2015
  9. N. Desai, P Rambhia and A. Gishto, Human embryonic stem cell cultivation: historical perspective and evolutionof xeno-free culture systems. Reproductive Biology and Endocrinology 13.1 (2015): 9.
  10. T. Yokobori et al., Intestinal epithelial culture under an air-liquid interface: a tool for studying human and mouse esophagi. Diseases of the Esophagus, doi: 10.1111/dote.12346. 2015
  11. L. Healy, L Ruban, Derivation of Induced Pluripotent Stem Cells, Atlas of Human Pluripotent Stem Cells in Culture, pp 149-165. Springer US 2015
  12. W. Siqin et al. Spider silk for xeno-free long-term self-renewal and differentiation of human pluripotent stem cells. Biomaterials 35.30 (2014): 8496-8502.
  13. G. Finesilver, M. Kahana, E. Mitrani. Kidney-Specific Micro-Scaffolds and Kidney Derived Serum FreeConditioned Media support in vitro Expansion, Differentiation, and Organization of Human Embryonic Stem Cells. Tissue Engineering Part C: Methods. -Not available-, ahead of print. doi:10.1089/ten.TEC.2013.0574.
  14. M. Amit, J. Itskovitz-Eldor. Atlas of Human Pluripotent Stem Cells: Derivation and Culturing. Stem Cell Biology and Regenerative Medicine, 2012
  15. R. Bergström, Xeno-free culture of human pluripotent stem cells, Methods Mol Biol. 2011;767:125-36
  16. J.Collins et al,. Highly Efficient Reprogramming to Pluripotency and Directed Differentiation of Human Cells withSynthetic Modified mRNA. Cell Stem Cell 7 (5): 618-630 (2010).
  17. K. Jacobs et al. Higher-Density Culture in Human Embryonic Stem Cells Results in DNA Damage and GenomeInstability. Stem Cell Reports: 6(3), pp 330–341, 2016

 Differentiation of Pluripotent Stem Cells

  1. K.M. Gray et al. Self-oligomerization regulates stability of Survival Motor Neuron (SMN) protein isoforms by sequestering an SCFSlmb degron. Molecular Biology of the Cell, 2017 mbc.E17-11-0627
  2. D.C. Wilkinson et. al. Development of a Three-Dimensional Bioengineering Technology to Generate Lung Tissue for Personalized Disease Modeling. Stem cells translational medicine, 6(2), 2017
  3. E. Welby et al. Isolation and Comparative Transcriptome Analysis of Human Fetal and iPSC-Derived Cone Photoreceptor Cells. Stem Cell Reports (2017), https://doi.org/10.1016/j.stemcr.2017.10.018
  4. R. De-Santis et. al. FUS Mutant Human Motoneurons Display Altered Transcriptome and microRNA Pathways with Implications for ALS Pathogenesis. Stem Cell Reports (2017), https://doi.org/10.1016/j.stemcr.2017.09.004
  5. R.A. Hazim et al. Differentiation of RPE cells from integration-free iPS cells and their cell biological characterization. Stem Cell Research & Therapy 2017
  6. S. Petrus-Reurer et al. Integration of Subretinal Suspension Transplants of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells in a Large-Eyed Model of Geographic Atrophy. Retinal Cell Biology, February 2017
  7. X. Yuan et al. A hypomorphic PIGA gene mutation causes severe defects in neuron development and susceptibility to complement-mediated toxicity in a human iPSC model, PLOS ONE, 2017
  8. Lenzi, J., et al. Differentiation of control and ALS mutant human iPSCs into functional skeletal muscle cells, a tool for the study of neuromuscolar diseases. Stem Cell Research: Volume 17, Issue 1, Pages 140–147, 2016.
  9. K. Alessandri et. al. A 3D printed microfluidic device for production of functionalized hydrogel microcapsules forculture and differentiation of human Neuronal Stem Cells (hNSC). Lab on a Chip: 16(9), 2016
  10. D. Voulgaris, Evaluation of Small Molecules for Neuroectoderm differentiation & patterning using Factorial Experimental Design. Master Thesis in Applied Physics, Department of Physics, Division of Biological Physics, Chalmers University of Technology, Göteborg, Sweden 2016
  11. P. Bergström et al. Amyloid precursor protein expression and processing are differentially regulated during cortical neuron differentiation, Scientific Reports, 2016
  12. Tieng, V. ae al. Elimination of proliferating cells from CNS grafts using a Ki67 promoter-driven thymidine kinase, Molecular Therapy — Methods & Clinical Development 6, Article number: 16069, 2016
  13. Brykczynska, U, et al. CGG Repeat-Induced FMR1 Silencing Depends on the Expansion Size in Human iPSCs and Neurons Carrying Unmethylated Full Mutations Stem Cell Reports, 2016
  14. Sellgren ,C.M. et al. Patient-specific models of microglia-mediated engulfment of synapses and neural progenitors Molecular Psychiatry, 2016
  15. Cosset, E. et al. Human tissue engineering allows the identification of active miRNA regulators of glioblastoma aggressiveness Biomaterials, 2016
  16. M. Di Salvio et al. Pur-alpha functionally interacts with FUS carrying ALS-associated mutations. Cell Death & Disease, 2015
  17. A. Reyes, et al. Xeno-Free and Defined Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells Functionally Integrate in a Large-Eyed Preclinical Model Plaza. Stem Cell Reports: Volume 6, Issue 1, p9–17, 2015
  18. Tieng, V. et al.Engineering of Midbrain Organoids Containing Long-Lived Dopaminergic Neurons. Stem Cells and Development. February 2014, 23(13): 1535-1547.
  19. A. J. Schwab, A.D. Ebert, Sensory Neurons Do Not Induce Motor Neuron Loss in a Human Stem Cell Model of SpinalMuscular Atrophy. PLoS One. 2014; 9(7): e103112
  20. H.X. Nguyen et al., Induction of early neural precursors and derivation of tripotent neural stem cells from humanpluripotent stem cells under xeno-free conditions. Journal of Comparative Neurology: Volume 522, Issue 12, pp 2767–2783, 2014
  21. A. Kurtz, A. Bosio, and S. Knoebel. Highly efficient differentiation of hPSC into hepatocyte-like cellsby selection of CXCR4 (CD184) definitive endoderm (DE) cells

Cardiomyocyte differentiation

  1. R. Ophir et al. Inflammation And Contractility Are Altered By Obstructive Sleep Apnea Children's Serum, In Human Embryonic Stem Cell Derived Cardiomyocytes. American Journal of Respiratory and Critical Care Medicine 2017
  2. J. Kristensson, Optimization of Growth Conditions for Expansion of Cardiac Stem Cells Resident in the Adult Human Heart. Master’s thesis in Biotechnology, Department of Physics, Division of Biological Physics, Chalmers University of Technology, Gothenburg, Sweden 2016
  3. S. Rajasingh et al. Generation of Functional Cardiomyocytes fro Efficiently Generated Human iPSCs and a Novel Method of Measuring Contractility. PloS one 10.8, 2015: e0134093
  4. L. Jacquet et al. Three Huntington’s Disease Specific Mutation-Carrying Human Embryonic Stem Cell Lines Have Stable Number of CAG Repeats upon In Vitro Differentiation into Cardiomyocytes. PloS one 10.5, 2015
  5. V. Bellamy et al., Long-term functional benefits of human embryonic stem cell-derived cardiac progenitors embedded into a fibrin scaffold, The Journal of Heart and Lung Transplantation, 2014, in press
  6. E. Di Pasquale et al. Generation of human cardiomyocytes: a differentiation protocol for feeder-free human induced pluripotent stem cells. JoVE (Journal of Visualized Experiments) 76 (2013): e50429-e50429
  7. P.W. Burridge and E.T Zambidis. Highly efficient directed differentiation of human induced pluripotent stem cells into cardiomyocytes. Pluripotent Stem Cells: Methods and Protocols. Methods in Molecular Biology, volume 997, pp 149-161, Humana Press, 2013.

Gene Editing

  1. Sweeney, CL et al.Targeted Repair of CYBB in X-CGD iPSCs Requires Retention of Intronic Sequences for Expression and Functional Correction,Molecular Therapy, 2017
  2. J. Lenzi et al., ALS mutant FUS proteins are recruited into stress granules in induced Pluripotent Stem Cells (iPSCs) derived motoneurons. Disease Models & Mechanisms: 8, 755-766, 2015
  3. T. Cerbini et al., Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targetedwith a GFP Gene into the AAVS1 Safe Harbor, JoVE (Journal of Visualized Experiments), 2015

 Induction of Pluripotency of hESC and iPSC

  1. X. Gao et. al. Comparative transcriptomic analysis of endothelial progenitor cells derived fro umbilical cord blood and adult peripheral blood: Implications for the generation of induced pluripotent stem cells. Stem Cell Research, 2017
  2. M.V. Krivega et al. Cyclin E1 plays a key role in balancing between totipotency and differentiation in human embryonic cells. Mol. Hum. Reprod, 2015
  3. S. Herz, Optimization of RNA-based transgene expression by targeting Protein Kinase R. Dissertation for the degree “Doctor rerum naturalium”, 2015
  4. S. Eminli-Meissner et al. A novel four transfection protocol for deriving iPS cell lines fro human blood- derivedendothelial progenitor cells (EPCs) and adult human dermal fibroblasts using a cocktail of non-modified reprogrammingand immune evasion mRNAs. Sientific Poster, REPROCELL, 2015
  5. M. Brouwer et al. Choices for Induction of Pluripotency: Recent Developments in Human Induced Pluripotent StemCell Reprogramming Strategies. Stem Cell Reviews and Reports: Volume 12, Issue 1, pp 54–72, 2015
  6. R. S. Song et al. Generation, Expansion, and Differentiation of Human Induced Pluripotent Stem Cells (hiPSCs) Derived fro the Umbilical Cords of Newborns. Current protocols in stem cell biology (2013): UNIT 1C.16.
  7. Warren, L., et al. Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell 7: 618-630, 2010
  8. S. Sugii et al., Human and mouse adipose-derived cells support feeder-independent induction of pluripotent stem cells. PNAS February 23, 2010 vol. 107 no. 8 3558-3563

Proteins and Antibodies Expression and Isolation

  1. D.R. Riordon and K.R. Boheler, Immunophenotyping of Live Human Pluripotent Stem Cells by Flow Cytometry. In: Boheler K., Gundry R. (eds) The Surfaceome. Methods in Molecular Biology, vol 1722. Humana Press, New York, NY, 2018
  2. N.Y. Thakar et al., TRAF2 recruitment via T61 in CD30 drives NFkB activation and enhances hESC survival andproliferation, Molecular Biology of the Cell: 26(5):993-1006 2015
  3. Abcam, Immunocytochemistry / Immunofluorescence abreview for Anti-Oct4 antibody - ChIP Grade. Abcam website

Clinical Applications- Derivation and Expansion of hESC and IPSC

  1. L. de Oñate et al. 2015. Research on Skeletal Muscle Diseases Using Pluripotent Stem Cells. DOI: 10.5772/60902
  2. P. Menasché et al., Towards a Clinical Use of Human Embryonic Stem Cell-Derived Cardiac Progenitors:A Translational Experience. European Heart Journal: Volume 36, Issue 12, pp 743-50, 2015
  3. P. Menasché et al. Human embryonic stem cell-derived cardiac progenitors for severe heart failure treatment: first clinical case report. European heart journal (2015): ehv189.
  4. T. Seki, K. Fukuda, Methods of induced pluripotent stem cells for clinical application, World Journal of Stem Cells: Volume 7, Issue 1, pp 116–125, 2015
  5. Y. Luo et al.,Stable Enhanced Green Fluorescent Protein Expression After Differentiation and Transplantation of Reporter Human Induced Pluripotent Stem Cells Generated by AAVS1 Transcription Activator-Like Effector Nucleases, STEM CELLS Translational Medicine: Volume 3, Issue 7, pp 821-35, 2014
  6. J. Durruthy-Durruthy et al. Rapid and Efficient Conversion of Integration-Free Human Induced Pluripotent Stem Cells to GMP-Grade Culture Conditions. PlOS one: http://dx.doi.org/10.1371/journal.pone.0094231, 2014
  7. H. Tateno et al. A medium hyperglycosylated podocalyxin enables noninvasive and quantitative detection of tumorigenic human pluripotent stem cells. Scientific Reports 4, Article number: 4069, 2014
  8. S. Abbasalizadeh, H. Baharvand. Technologies progress and challenges towards cGMP manufacturing of human pluripotent stem cells based therapeutic products for allogeneic and autologous cell therapies. Biotechnology Advances: Volume 31, Issue 8, pp 1600-23, 2013.
  9. J. P. Awe et al. Generation and characterization of transgene-free human induced pluripotent stem cells and conversion to putative clinical-grade status. Stem Cell Research & Therapy, 2013, 4:87
  10. O. Hovatta. Infectious problems associated with transplantation of cells differentiated fro pluripotent stem cells. Seminars in Immunopathology: Volume 33, Issue 6, pp 627-30, April 2011
  11. S. Ström. Optimisation of human embryonic stem cell derivation and culture – towards clinical quality. Karolinska Institutet, Stockholm, Sweden, 2010

Drug Screening

  1. Z. Ye et al., Differential sensitivity to JAK inhibitory drugs by isogenic human erythroblasts and hematopoieticprogenitors

Animal Models

  1. W. Zhang, et al. Distinct MicroRNA Expression Signatures of Porcine Induced Pluripotent Stem Cells under Mouse and Human ESC Culture Conditions. PLOS ONE, 2016

Patents

  1. J. Mata-fink et al. Methods and Compositions For Immunomodulation. US Patent Application 20170020926, 2017
  2. Bailly, J., et al. Method for differentiation of pluripotent stem cells into multi-competent renal precursors. US Patent 20,160,145,578, 2016
  3. K. Tryggvason et al. Differentiation of pluripotent stem cells and cardiac progenitor cells into striated cardiomyocyte fibers using laminins LN-511, LN-521 and LN-221. US Patent Application 20160122717, 2016
  4. D. Keefe et al. A Method for a Single Cell Analysis of Telomere Length. US Patent Application 20160032360, 2016
  5. Dietrich M. EGLI, Methods for making and using modified oocytes . US Patent Application 20140308257, 2016
  6. M. Mikkola et. al. A Method for Generating Induced Pluripotent Stem Cells. US Patent Application 20160068818, 2016
  7. H Tateno et al. Undifferentiated Cell Detection Method and Complex Carbohydrate Detection Method. US Patent Application 20150204870, 2015
  8. O. Hovatta, K. Tryggvason, Methods of Producing RPE Cells. US Patent Application 20150299653, 2015
  9. A. De Fougerolles, S.M. Elbashir, Delivery and Formulation of Engineered Nucleic Acids, US Patent Application 20150017211, 2015
  10. Sahin, U., et al. Diagnosis and therapy of cancer involving cancer stem cells. US Patent Application 20150314018, 2015
  11. U. Sahin et al. Method for Cellular RNA Expression. US Patent Application 20150314018, 2015
  12. M.F Yanik, M. Angel. Methods for Transfecting Cells With Nucleic Acids. US Patent Application 20140073053, 2014
  13. C.E Buensuceso et al., Induced Pluripotent Stem Cells Prepared fro Human Kidney-Derived Cells. US Patent Application 20140073049, 2014
  14. C. Buensuceso et al., Induced Pluripotent Stem Cells fro Human Umbilical Cord Tissue-Derived Cells. US Patent Application 20130157365, 2013
  15. A. De Fougerolles, S.M. Elbashir, J.P. Schrum. Modified Polynucleotided for the Production of Factor IX. US Patent Application 20130245103, 2013
  16. M. Amit, J. Itskovitz-eldor. Novel Methods and Culture Media for Culturing Pluripotent Stem Cells. US Patent Application 20130236961, 2013
  17. E. Zambidis, P Burridge. Compositions and Methods of Generating a Differentiated Mesodermal Cell. US Patent Application 20130136721, 2013.

Reviews

There are no reviews yet.


Be the first to review “Nutristem hPSC XF Medium”