A comparative study of the fibroblasts growth curve isolated from neck and distal limb of horse skin

Fibroblasts are one of the important cells in wound healing. These cells create a proper bed for keratinocytes migration and wound contraction.Wound healing in distal limb of horses has complications, such as formation of exuberant granulation tissue (EGT). The main factor in this problem is overgrowing of fibroblasts. The purpose of the present study was to compare fibroblast growth curve in isolated skin from horses’ neck and distal limb. 5 horses with normal hematological and clinical signs were selected. Two samples of full thickness of skin were taken from the neck and lateral metacarpal region of each horse asseptically. Then the samples were washed with PBS minced and placed in ventilated flask 25 cm2. After attaching samples to flask, 5 ml culture medium(RPMI-1640 with 10% FBS) were added and the flask was placed in an incubator at 37°c in 5% CO2. After leaving a sufficient number of cells from tissues adhered to the bottom of the flask, the cells were passaged to a new ventilated flask. After growth and proliferation of cells, they were passaged again and a suspension of cells in culture medium (10000 cells/ml) was maked. To each cell of a 24-well plate, one ml of this suspension was added. After 48 hour, cells of 3 well were detached with tripsin daily, counted and viability determinted within 8 days. There was no significant difference between viable cells number but there was significant difference in viability percent of cells in neck and distal limb. The mean of population doubling time (PDT) for fibroblasts of neck is 31.73 hours and for fibroblasts of distal limb is 26.4 hours. This difference was not significant. With regard to different viability percentage, it seems that the appoptosis in fibroblasts of neck skin is more regular than distal limb skin.