Dissociation solution for human ES/iPS cells

Product Classification : hES/iPSC Culture Media / ReagentsResearch Theme : ES/iPS cells

Dissociation solution for human ES/iPS cells

Dissociation Solution for Human ES/iPS Cells (CTK solution)

  • Gentle detachment of the colonies without using scrapers*
  • 10 times more viability compared to trypsin
  • Long term culture without karyotype abnormality
  • For both of feeder-dependent and feeder-free culture
  • Each lot is functionally tested

(Storage at -20 degC)

Overview

Relative survwal rate 1:10

ReproCELL's dissociation solution is well known as CTK (Collagenase Type IV, Trypsin, KSR) solutions among researchers, which was developed by Prof. Norio Nakatsuji, Kyoto University Institute for Integrated Cell Material Sciences (iCeMS) founding director and Assoc. Prof. Hirofumi Suemori, Kyoto University Institute for Frontier Medical Sciences. Currently, "dissociation solution for human ES cells and iPS cells" is widely used to dissociate cells among stem cell researchers.

When passaging of human ES cells and iPS cells, it is crucial to maintain colony without preparing single cell suspension. For example, when tripcin-EDTA solution is used to detach and passage human ES cells or monkey ES cells, these cell viability greatly decreases. On the other hand, ReproCELL'S dissociation solution enhanced cell viability at passaging close to 10-folds higher than that of using tripcin-EDTA solution.

Another benefit of using ReproCELL's dissociation solution is that colony size will be optimized for passaging only by adding RepoCELL's dissociation solution. (Please see the pictures below) In fact you do not have to scrape cells to dissociate. Also, chromosomal abnormality have not been observed during human and cynomolgus monkey ES cell culture for more than 3 years.

Figure: Passaging using Dissociation solution for human ES/iPS cells (RCHETP002)

[movie] Detachment of an iPSC colony by Dissociation Solution (RCHETP002)

Protocol

Data Sheet

Safety Data Sheet

Instruction Movies

The culture procedures for human iPS cells

Instruction movies for human iPS cell culture procedure with ReproCELL's culture reagents.

Chapter 1: Human iPSC colony morphology

Morphology of human iPS cell colonies is important to judge the condition of the culture. Colonies in a good condition show high cell density inside.

Chapter 2: Seeding feeder cells

The plating density of feeder cells is important for human iPS cell culture.

Chapter 3: Passaging human iPS cells

Dissociation Solution for human ES/iPS Cells (CTK solution) makes it possible to passage human iPS cells very easily and successfully with excellent cell viability. Colonies can be divided to appropriate size for passaging just by incubation and pipetting.

Chapter 4: Freezing and thawing human iPS cells

It is possible to cryopreserve human iPS cells at high cell viability by using Freezing Medium for human ES/iPS Cells. This method ulitizes vitrification which minimizes the damage to the cryopreserved cells. It is important to freeze and thaw the cells quickly.

Publications

  • Morizane R, et al. “Nephron organoids derived from human pluripotent stem cells model kidney development and injury.” Nat Biotechnol. 2015 Oct 12. doi: 10.1038/nbt.3392.
  • Kumazaki, Tsutomu, et al. "Re‐emergence of undifferentiated cells from transplants of human induced pluripotent stem cells as a possible risk factor of tumourigenesis." Cell Biology International Reports (2014).
  • Delacote, Fabien, et al. "High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes." PloS one 8.1 (2013): e53217.
  • Miyazaki, Takamichi, Norio Nakatsuji, and Hirofumi Suemori. "Optimization of slow cooling cryopreservation for human pluripotent stem cells." genesis (2013).
  • Razak, Siti Razila Abdul, et al. "Profiling of MicroRNA in Human and Mouse ES and iPS Cells Reveals Overlapping but Distinct MicroRNA Expression Patterns." PloS one 8.9 (2013): e73532.
  • Kuroda, Takuya, et al. "Highly sensitive in vitro methods for detection of residual undifferentiated cells in retinal pigment epithelial cells derived from human iPS cells." PloS one 7.5 (2012): e37342.
  • Kobayashi, Hideyuki. "Pluripotent Stem Cells Induced from Testicular Tissue of a Man with Klinefelter Syndrome (47, XXY) by Four Transcription Factors (OCT4, SOX2, KLF4, and C-MYC)." (2011).
  • Kumazaki, Tsutomu, et al. "Establishment of human induced pluripotent stem cell lines from normal fibroblast TIG-1." Human cell 24.2 (2011): 96-103.
  • Shirasawa, Sakiko, et al. "Pancreatic exocrine enzyme-producing cell differentiation via embryoid bodies from human embryonic stem cells." Biochemical and biophysical research communications 410.3 (2011): 608-613.
  • Oka, Y., et al. "293FT cells transduced with four transcription factors (OCT4, SOX2, NANOG, and LIN28) generate aberrant ES-like cells." J Stem cell Regenerative Med 3 (2010): 149-56.
  • Lee, Tae-Hee, et al. "Functional recapitulation of smooth muscle cells via induced pluripotent stem cells from human aortic smooth muscle cells."Circulation research 106.1 (2010): 120-128.
  • Shimoji, Kenichiro, et al. "G-CSF promotes the proliferation of developing cardiomyocytes in vivo and in derivation from ESCs and iPSCs." Cell Stem Cell6.3 (2010): 227-237.
  • Shimoji, Kenichiro, et al. "G-CSF promotes the proliferation of developing cardiomyocytes in vivo and in derivation from ESCs and iPSCs." Cell Stem Cell6.3 (2010): 227-237.
  • Yoshie, Susumu, et al. "Bone morphogenetic protein-4 promotes induction of." J Cell Sci 122.17 (2009): 3070-3082.

Product list

Catalog No. Product Price
RCHETP002 Dissociation solution for human ES/iPS cells (30mL) Ask

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