Biological properties of C57BL/6 mouse embryonic fibroblasts and preparation of feeder layers / 中国组织工程研究

ABSTRACT

BACKGROUND:Kunming mouse embryonic fibroblasts are the most common feeder layers at present, and there are rare reports addressing C57BL/6 mouse embryonic fibroblasts as feeder layers. OBJECTIVE:To separate and culture C57BL/6 mouse embryonic fibroblasts in vitro, and produce feeder layers to enlarge the resources of mouse embryonic fibroblasts. METHODS:C57BL/6 mouse embryonic fibroblasts were isolated and cultured by trypsin digestion method in vitro. The biological characteristics and growth rule of the fibroblasts were investigated, then the feeder layers for the cel culture were produced. The growth of cel colonies on the prepared feeder layer was tested. RESULTS AND CONCLUSION:C57BL/6 mouse embryonic fibroblasts grew wel with a large amount, by trypsin digestion method at different concentrations. There was no significance in the survival rate after cryopreservation for 1 week, 2 weeks, 1 month, 3 months and 6 months. The cel s were proliferative from the second to fifth passage and declined sharply after the sixth passage. The planted mouse embryonic fibroblasts feeder layers had a high activity within 3 days, but got a sharp decline after 4 days. So it is best to use C57BL/6 mouse embryonic fibroblast feeder layers within 3 days after they’re inactivated. C57BL/6 mouse embryonic fibroblast feeder layer can support embryonic stem cel s and induce pluripotent stem cel s to grow as Kunming mouse embryonic fibroblasts.