CryoDefend-Stem Cells (5 x 10 mL)

Human Embryonic Stem Cell Viability Following Cryopreservation in CryoDefend® Stem Cells Media or Cryopreservation Media from Two Competitors.  Stem Cells Media or Cryopreservation Media from Two Competitors.BG01V human embryonic stem cells were frozen in cryopreservation media from two different competitors or CryoDefend^®Stem Cells Media (Catalog # CCM018) at 1 x 10⁶cells/cryovial and stored in liquid nitrogen for one week. BG01V human embryonic stem cells were then thawed, counted (blue bars), and resuspended in Mouse Embryonic Fibroblast (MEF) Conditioned Media (Catalog # AR005) containing Recombinant Human FGF basic (Catalog # 4114-TC). The resuspended cells were plated into one well of a 6-well plate and after four days in culture, the cells were harvested and counted (red bars). The error bars indicate the standard deviation of triplicate samples.

Morphology of Human Embryonic Stem Cells Following Cryopreservation in CryoDefend® Stem Cells Media, Conventional Freezing Media, or Freezing Media from a Competitor. BG01V human embryonic stem cells were frozen in CryoDefend^®Stem Cells Media (Catalog # CCM018), conventional freezing media (90% FBS/10% DMSO) or cryopreservation media from a competitor. After thawing, the cells were cultured for 3 days in Mouse Embryonic Fibroblast Conditioned Media (Catalog # AR005) containing FGF basic (Catalog # 4114-TC) and imaged using brightfield microscopy.

Expression of Pluripotency Markers in BG01V Human Embryonic Stem Cells Frozen in CryoDefend® Stem Cells Media or Cryopreservation Media from Two Different Competitors. Cells that were cryopreserved in CryoDefend^®Stem Cells Media (Catalog # CCM018) or cryopreservation media from one of two competitors CryoDefend^®Stem Cells Media (Catalog # CCM018) were thawed and cultured for four days in Mouse Embryonic Fibroblast Conditioned Media (Catalog # AR005). The cells were then assessed for pluripotency marker expression by flow cytometry. The cells were stained with APC-conjugated Mouse Anti-Human Oct-4A (Catalog # IC6344A) and PE-conjugated Mouse Anti-Human/Mouse SSEA-4 (Catalog # FAB1435P). The quadrants were set based on isotype controls. The percent of double positive cells are indicated in the upper right quadrant.

Human Embryonic Stem Cells Maintain Pluripotency Following Cryopreservation in CryoDefend® Stem Cells Media. BG01V human embryonic stem cells that were frozen in CryoDefend^®Stem Cells Media (Catalog # CCM018) were thawed and cultured for four days before the cells were replated for differentiation into each of the three germ layers according to the insert instructions for the Human Pluripotent Stem Cell Functional ID Kit (Catalog # SC027). Differentiated cells were then stained for markers of each germ layer as indicated in the images using the fluorochrome-conjugated antibodies included in the Human Three-Germ Layer 3-Color Immunocytochemistry Kit (Catalog # SC022). The nuclei were counterstained with DAPI (blue).

Recovery and Marker Expression of Rat Mesenchymal Stem Cells Cryopreserved in CryoDefend® Stem Cells Media or Conventional Freezing Media.  Rat Mesenchymal Stem Cells (Catalog # PSC003) were cryopreserved in CryoDefend^®Stem Cells Media (Catalog # CCM018) or conventional freezing media (95% FBS/5% DMSO) at 0.7x10⁶cells/vial. The cryopreserved cells were thawed and cultured for four days before assessing cell recovery and MSC marker expression. A) The number of cryopreserved cells recovered after four days in culture. B) Expression of positive MSC marker CD90 and negative MSC marker CD45 after the thawed cryopreserved cells were cultured for four days. CD90 was detected with a RPE-conjugated Anti-Rat CD90/Thy1 Antibody and CD45 was detected with 687-conjugated Anti-Rat CD45 Antibody.

Rat Cortical Stem Cell Viability Following Cryopreservation in Control Cyropreservation Media or CryoDefend® Stem Cells Media.  Rat cortical stem cells (Catalog # NSC001) were frozen in control cryopreservation media (DMEM/F12 supplemented with N2-MAX Media Supplement,10% BSA, and 10% DMSO) or in CryoDefend^®Stem Cells Media (Catalog # CCM018) at 1 x 10⁶cells per cryovial and stored in liquid nitrogen for one week. After thawing in DMEM/F12 containing N2-MAX Media Supplement (Catalog # AR009) and 20 ng/mL of FGF basic (Catalog # 4114-TC), the cells were counted (light blue bars) and plated onto Poly-L-ornithine/Fibronectin-coated plates. The cells were cultured for four days prior to passage (Passage 1) at which time the cells were counted (dark blue bars). The error bars indicate the standard deviation of triplicate samples.