[Long-term in vitro culture of the fibroblasts from the deep partial thickness burn wound in burn patients].
OBJECTIVE
To establish a long-term in vitro culture of the fibroblasts obtained from burn wounds.


METHODS
Skin samples were harvested from normal volunteers and the deep partial thickness burn wound in burn patients on the 5th, 10th, 21st, 28th and 35th postburn days (PBDs). The non-dermal tissue was removed from the samples and primed by chlorhexidine solution in concentration of 2.5 g/L. The skin sample was then digested by trypsin-EDTA in concentration of 1.25 g/L and was centrifuged before the cells were harvested and cultured. When the cells grew nearly to form sheet, multiple passage culture, freezing storage and revivification were carried out with routine methods. The cell morphology was continuously observed during the culture. And the cell doubling time was calculated.


RESULTS
The wound-origin fibroblasts exhibited higher purity and better activity. The cellular growth features and gross morphology kept stable during primary and secondary culture, and during freezing storage and after revivification. The cells kept their activity above 80% of their original after many times of revivification.


CONCLUSIONS
The establishment of the in vitro culture of fibroblasts from burn wounds might be useful in the exploration of the pathogenesis and therapeutic measures of scars.
Li
Yong-Lin
YL
Department of Burns, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, P.R. China.
Chen
Bi
B
Tang
Chao-Wu
CW
Xu
Ming-Da
MD
D002056:Burns / Q000378:metabolism / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D005169:Factor VIII / Q000032:analysis; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D013997:Time Factors
D005169:Factor VIII