Human Umbilical Cord Blood-Derived Serum for Culturing the Supportive Feeder Cells of Human Pluripotent Stem Cell Lines

Figure 1

Effect of serum supplementation on the growth kinetics of human foreskin fibroblasts. Human foreskin fibroblasts were enzymatically dissociated and plated onto culture dishes. HFFs were cultured in the media containing human cord blood-derived serum (hUCS; HFF-hUCS), fetal bovine serum (FBS; HFF-FBS), knockout serum replacement (KSR) or without serum supplementation. Dissociated cells were observed for attachment behavior and growth kinetics. Differences in cell attachment between HFF-hUCS and HFF-FBS were observed. At 5 hours after plating, nearly all of the dissociated HFF-FBS were attached to the surface of the culture dish and exhibited a fibroblastic-like morphology, while most of HFF-hUCS remained suspended in the culture medium. At 24 hours after plating, all HFF-FBS and HFF-hUCS were attached to the culture dishes and displayed typical fibroblast cell morphology (a). Similar proliferation patterns between HFF-hUCS and HFF-FBS were observed. HFF-hUCS and HFF-FBS progressively proliferated and reached confluency at 3 to 4 days after seeding. The proliferation behavior at early passage (p4 + 1) was similar to that at intermediate passage (p4 + 10) (b). In contrast to HFF-hUCS and HFF-FBS, HFFs cultured in the medium containing 10% KSR and without serum supplementation could not reach the confluency even being cultured for 7 days (c). HFF = human foreskin fibroblasts, FBS = fetal bovine serum, hUCS = human cord blood-derived serum, KSR = knockout serum replacement, and p = passage number. Scale bars = 200 μm.

(a)

(b)

(c)