NutriStem® hPSC XF Medium

Defined, xeno-free, serum-free medium
Designed for optimal growth and expansion of human iPS and hES cells
Name SKU Size Price Qty
NutriStem® hPSC XF Culture Medium 05-100-1A 500 mL
$240.00
NutriStem® hPSC XF Culture Medium 05-100-1B 100 mL
$65.00
NutriStem® hPSC XF Culture Medium (Modified, GF-free) 06-5100-01-1A 500 mL
$215.00

Description

Details

Product Overview

NutriStem® hPSC XF Medium is a widely published, 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. Protocols have been established around the world for applications ranging from derivation to differentiation. NutriStem® hPSC XF Medium offers the ability to culture cells in a completely xeno-free medium without the need for high levels of basic FGF and other stimulatory growth factors and cytokines. In addition, superior cell attachment and proliferation is observed with NutriStem® hPSC XF Medium aid high-throughput screening applications. NutriStem® hPSC XF Medium exhibits a consistent media performance and predictable cellular behavior derived from a defined xeno-free formulation as well as increased reproducibility shown in long-term growth of over 50 passages.

Low-protein formulation that contains stable L-alanyl-L-glutamine and HSA.

Features

  • Defined, serum-free, and xeno-free 
  • Flexible and compatible with multiple matrices (including vitronectin, laminin, and Matrigel)
  • Amenable to weekend-free culture
  • Drug Master File (DMF) available, produced under cGMP
  • Enables efficient expansion and growth of hES and hiPS cells in feeder-free culture systems
  • Extensively tested and widely used on multiple hES and iPS cell lines, including H1, H9.2, I6, I3.2, and CL1
hESC were cultured in NutriStem® hPSC XF


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.

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).

Taratoma formation

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).

Specifications

Specifications

Form Liquid
Brand NutriStem®
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.
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 are recombinant laminin, such as LaminStem(R) 521 (BI Cat. No. 05-753-1F), and recombinant vitronectin, which have 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.

References

references

NutriStem® and Cardiomyocytes

  1. 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
  2. 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.
  3. L. Jacquet et al. Three Huntington’s Disease Specific Mutation-Carrying Human Embryonic Stem Cell Lines HaveStable Number of CAG Repeats upon In Vitro Differentiation into Cardiomyocytes. PloS one 10.5, 2015
  4. S. Rajasingh et al. Generation of Functional Cardiomyocytes from Efficiently Generated Human iPSCs and a NovelMethod of Measuring Contractility. PloS one 10.8, 2015: e0134093
  5. 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.
  6. E. Di Pasquale et al. Generation of human cardiomyocytes: a differentiation protocol from feeder-free humaninduced pluripotent stem cells. JoVE (Journal of Visualized Experiments) 76 (2013): e50429-e50429
  7. G. Földes and M. Mioulane. High-content imaging and analysis of pluripotent stem cell-derived cardiomyocytes.Imaging and Tracking Stem Cells. Humana Press, 2013.
  8. P.W. Burridge and E.T Zambidis. Highly efficient directed differentiation of human induced pluripotent stem cellsinto cardiomyocytes. Pluripotent Stem Cells: Methods and Protocols. Methods in Molecular Biology, volume 997, pp 149-161, Humana Press, 2013.
  9. R. S. Song et al. Generation, Expansion, and Differentiation of Human Induced Pluripotent Stem Cells (hiPSCs)Derived From the Umbilical Cords of Newborns. Current protocols in stem cell biology (2013): UNIT 1C.16.
  10. 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


  11. Cancer Stem Cells

  12. Y Qin, et al. Laminins and cancer stem cells: partners in crime? Seminars in Cancer Biology, 2016


  13. Differentiation of Pluripotent Stem Cells

  14. P. Bergström et al. Amyloid precursor protein expression and processing are differentially regulated during cortical neuron differentiation, Scientific Reports, 2016
  15. 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
  16. 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
  17. 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
  18. Sellgren ,C.M. et al. Patient-specific models of microglia-mediated engulfment of synapses and neural progenitors Molecular Psychiatry, 2016
  19. Bailly, J., et al. Method for differentiation of pluripotent stem cells into multi-competent renal precursors. US Patent 20,160,145,578, 2016
  20. 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.
  21. 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
  22. 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
  23. 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
  24. S. Rajasingh et al. Generation of Functional Cardiomyocytes from Efficiently Generated Human iPSCs and a NovelMethod of Measuring Contractility. PloS one 10.8, 2015: e0134093


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

  26. Baker, D. et al. Detecting Genetic Mosaicism in Cultures of Human Pluripotent Stem Cells Stem Cell Reports, 2016
  27. 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)
  28. W. Zhang, et al. Distinct MicroRNA Expression Signatures of Porcine Induced Pluripotent Stem Cells under Mouse and Human ESC Culture Conditions. PLOS ONE, 2016
  29. M. Mikkola et. al. A Method for Generating Induced Pluripotent Stem Cells. US Patent Application 20160068818, 2016
  30. 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
  31. 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
  32. S. Eminli-Meissner et al. A novel four transfection protocol for deriving iPS cell lines from 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
  33. S. Wu et al. Efficient passage of human pluripotent stem cells on spider silk matrices under xeno-free conditions. Cellular and Molecular Life Sciences: 73(7):1479-88, 2015
  34. 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
  35. H Tateno et al. Undifferentiated Cell Detection Method And Complex Carbohydrate Detection Method. US Patent Application 20150204870, 2015
  36. S. Herz, Optimization of RNA-based transgene expression by targeting Protein Kinase R. Dissertation for the degree “Doctor rerum naturalium”, 2015
  37. 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
  38. 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.
  39. 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
  40. 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
  41. L. Healy, L Ruban, Derivation of Induced Pluripotent Stem Cells, Atlas of Human Pluripotent Stem Cells in Culture, pp 149-165. Springer US 2015
  42. 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
  43. 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
  44. 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
  45. 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.
  46. M. Amit, J. Itskovitz-eldor. Novel Methods and Culture Media for Culturing Pluripotent Stem Cells. US Patent Application 20130236961, 2013
  47. M. Amit, J. Itskovitz-Eldor. Atlas of Human Pluripotent Stem Cells: Derivation and Culturing. Stem Cell Biology and Regenerative Medicine, 2012
  48. R. Bergström, Xeno-free culture of human pluripotent stem cells, Methods Mol Biol. 2011;767:125-36
  49. 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
  50. 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).

Induction of Pluripotency of hESC and iPSC

  1. S. Eminli-Meissner et al. A novel four transfection protocol for deriving iPS cell lines from 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
  2. Sahin, U., et al. Diagnosis and therapy of cancer involving cancer stem cells. US Patent Application 20150314018, 2015
  3. 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
  4. 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


  5. iPSC Derivation Patents

  6. U. Sahin et al. Method for Cellular RNA Expression. US Patent Application 20150314018, 2015
  7. M.F Yanik, M. Angel. Methods for Transfecting Cells With Nucleic Acids. US Patent Application 20140073053, 2014
  8. C.E Buensuceso et al., Induced Pluripotent Stem Cells Prepared from Human Kidney-Derived Cells. US Patent Application 20140073049, 2014
  9. C. Buensuceso et al., Induced Pluripotent Stem Cells from Human Umbilical Cord Tissue-Derived Cells. US Patent Application 20130157365, 2013
  10. A. De Fougerolles, S.M. Elbashir, J.P. Schrum. Modified Polynucleotided for the Production of Factor IX. US Patent Application 20130245103, 2013


  11. Different Basement Matrices

  12. Cosset, E. et al. Human tissue engineering allows the identification of active miRNA regulators of glioblastoma aggressiveness Biomaterials, 2016
  13. Wilkinson, D.C, et al. Development of a Three-Dimensional Bioengineering Technology to Generate Lung Tissue for Personalized Disease Modeling Stem Cells Translational Medicine, 2016
  14. O. Simonson. Use of Genes and Cells in Regenerative Medicine. Karolinska Institutet, 2015
  15. Nacalai USA Inc. Vitronectin-398™ (Xeno-free). Nacalai USA website
  16. S. Rodin et al., Monolayer culturing and cloning of human pluripotent stem cells on laminin-521–based matrices under xeno-free and chemically defined conditions. Nature Protocols 9, 2354–2368 (2014) doi:10.1038/ nprot.2014.159
  17. Rodin S, et al. Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment. Nat Commun. 5:3195. doi: 10.1038/ncomms4195, 2014
  18. StemAdhere™ Defined Matrix for hPSC. Primorigen Biosciences website.
  19. I. lenz et al., Automated 3D Culture to Undifferentiated hESC. (Scientific Poster)
  20. J.L. Weber et al,. The Corning® Synthemax™ Surface: A Synthetic, Xeno-Free Surface for Long-Term Self-Renewal of Human Embryonic Stem Cells in Defined Media. presented in 2010 world stem cell summit


  21. Proteins and Antibodies Expression and Isolation

  22. J. Mata-fink et al. Methods and Compositions For Immunomodulation. US Patent Application 20170020926, 2017
  23. Abcam, Immunocytochemistry / Immunofluorescence abreview for Anti-Oct4 antibody - ChIP Grade. Abcam website


  24. Clinical Applications- Derivation and Expansion of hESC and IPSC

  25. 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
  26. D. Keefe et al. A Method for a Single Cell Analysis of Telomere Length. US Patent Application 20160032360, 2016
  27. M. Di Salvio et al. 2015. Pur-alpha functionally interacts with FUS carrying ALS-associated mutations. Cell Death & Disease.
  28. L. de Oñate et al. 2015. Research on Skeletal Muscle Diseases Using Pluripotent Stem Cells. DOI: 10.5772/60902
  29. 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
  30. 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
  31. 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.
  32. 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
  33. J. Durruthy-Durruthy et al. 2014. 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
  34. H. Tateno et al. 2014. A medium hyperglycosylated podocalyxin enables noninvasive and quantitative detection of tumorigenic human pluripotent stem cells. Scientific Reports 4, Article number: 4069.
  35. 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
  36. Hovatta, Outi. Infectious problems associated with transplantation of cells differentiated from pluripotent stem cells. Seminars in Immunopathology: Volume 33, Issue 6, pp 627-30, April 2011
  37. Susanne Ström. Optimisation of human embryonic stem cell derivation and culture – towards clinical quality. Karolinska Institutet, Stockholm, Sweden, 2010.


  38. Patents

  39. Dietrich M. EGLI, Methods for making and using modified oocytes . US Patent Application 20140308257, 2016
  40. O. Hovatta, K. Tryggvason, Methods of Producing RPE Cells. US Patent Application 20150299653, 2015
  41. A. De Fougerolles, S.M. Elbashir, Delivery and Formulation of Engineered Nucleic Acids, US Patent Application 20150017211, 2015
  42. E. Zambidis, P Burridge. Compositions and Methods of Generating a Differentiated Mesodermal Cell. US Patent Application 20130136721, 2013.
  43. A. Kurtz, A. Bosio, and S. Knoebel. Highly efficient differentiation of hPSC into hepatocyte-like cellsby selection of CXCR4 (CD184) definitive endoderm (DE) cells


  44. Drug Screening

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

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