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Cell Reservoir One

Serum-Free Cell Freezing Medium

Cell Reservoir One is a serum-free cell culture freezing medium, which contains a water-soluble glycoprotein SERICIN isolated from the silkworm cocoon as a major constituent. SERICIN shows the same high efficacy of cryopreservation as FBS, and reduces the cell toxicity of DMSO. As DMSO is known to have adverse effects on cellular functions, especially Embryonic Stem Cells, Cell Reservoir One is available both with and without DMSO.

Application 1

Mouse ES cells:Viability after thawing

Jurkat: Human T Cell Lymphoblast-like Cell Line, mES: Mouse ES Cell, Freezing condition: -80°C, 1 week. The viability of Mouse ES cells in Cell Reservoir One is much higher than the one in FBS with 10%DMSO.

Mouse ES cells: Proliferative ability and undifferentiated state (3 days after thawing)

Freezing condition: -80°C, 1 week, Cell: Mouse ES Cells

Mouse ES cells: Differentiation into Beating cardiomyocytes

Freezing condition: -80°C, 1 week, Cell: Mouse ES Cells, Cell Reservoir One maintains high proliferation differentiation ability after thawing.

Mouse ES cells: Freezing media affect the differentiation

Freezing condition: -80°C, 1 week, Cell: Mouse ES Cells, Cultured with 10% of freezing media without Centrifugation. Cell Reservoir One maintains high differentiation ability without centrifugation after thawing cells.

Application 2

Human Mesenchymal Stem Cell (hMSC): Bone differentiation

Cell Reservoir One maintains bone differentiation ability of hMSC. Freezing condition: -80°C, 1 month, Condition: (Left)Alizarin Red staining after 4-weeks differentiation induction of hMSC. (Right)Quantification of calcium deposition after differentiation induction

Application 3

Pancreatic islet transplantation model

Bioartificial liver model

Cell viabilities in Cell Reservoir One with DMSO and without DMSO

Animal SpeciesCell SpeciesDerivationFreezing PeriodViability rate (%)
with DMSOwithout DMSO
Human Jurkat Blood 1 year 90 85
Human HL-60 Blood 1 year 90 80
Human WIL2-NS Spleen 1 year 90 70
Human MSC Bone Marrow 1 year 90 70
Human HeLa Uterine Cervix 1 year 90 80
Human HepG2 Liver 1 year 85 70
Human NHDF Normal Dermis 1 year 90 75
Mouse KUSA-A1 Bone Marrow 9 months 90 80
Mouse 2E3-O Hybridoma 1 year 90 80
Mouse P3U1 Bone Marrow 1 year 90 80
Mouse OP9 Skullcap 10 months 95 85
Mouse ES Embryo 1 year 80 90
Rat PC-12 Adrenal Gland 1 year 90 70
Hamster CHO Ovary 1 year 95 90
Rabbit RC4 Cornea 1 year 90 70
Insect Sf-9 Ovary 1 year 75 70

Protocol

[Freezing]

  1. Collect culture cells at their logarithmic growth phase.
  2. Suspend 5x105 -1x107 cells in 1 ml of Cell Reservoir One, and dispense the cell suspension in cryogenic vials.
  3. Freeze the cells at -80°C without preliminary freezing. The cells frozen at -80°C can be subsequently preserved in liquid nitrogen.

[Recovery]

  1. Remove the frozen vial from cryopreservation storage and rapidly thaw it in a 37°C water bath.
  2. Gently dilute the cells with cell culture medium by mixing.
  3. Centrifuge the cell suspension and remove the supernatant*, and resuspend the cells in culture medium.
    *Centrifugation step may be omitted in the case Cell Reservoir One without DMSO is used for ES cells.

Reference

  1. Aoki, Hiromasa, et al. "Efficient differentiation and purification of human induced pluripotent stem cell-derived endothelial progenitor cells and expansion with the use of inhibitors of ROCK, TGF-β, and GSK3β." Heliyon 6.3 (2020): e03493.
  2. Sawada, Ryoichi, et al. "High blood levels of soluble OX40 (CD134), an immune costimulatory molecule, indicate reduced survival in patients with advanced colorectal cancer." Oncology reports 42.5 (2019): 2057-2064.

Ordering Information

Product Cat.No. Storage PKG Size Price(US$)  
Cell Reservoir One (with DMSO) 07485-86 4°C 10 ml 24.00 Buy
Cell Reservoir One (with DMSO) 07485-44 4°C 100 ml 116.00 Buy
Cell Reservoir One (without DMSO) 07579-37 4°C 10 ml 29.00 Buy
Cell Reservoir One (without DMSO) 07579-24 4°C 100 ml 116.00 Buy