title: A modified culture strategy of human keratinocytes to shorten the primary culture time.
abstract: Shortening time from biopsy to graft preparation is crucial for the coverage of extensive burn wound in major burn patients. This study was aimed to shorten the culture time of keratinocytes by means of one modified cell culture method. Three factors (temperature of dispase, area of skin, and age of patient) that might have impacts on cell behavior were evaluated. The results showed that cells isolated with dispase at 37°C exhibited an improved cell activity than being isolated at 4°C. The time that cells reached subconfluence with various skin areas was in order of 3 cm(2)  < 2 cm(2)  < 1 cm(2) (120.07 ± 5.03 h, 141.33 ± 4.16 h, and 193.33 ± 6.11 h, respectively). Furthermore, compared with the young and middle-aged groups, proliferation of keratinocytes isolated from children skin tissue was better. These results indicated that 3 cm(2) skin biopsy from a young patient, isolated with dispase at 37°C, was a promising strategy to shorten the cultivation of keratinocytes.
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Le
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Zhan Jun
authors.initials: ZJ
authors.lastname: Lei
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Cong Ying
authors.initials: CY
authors.lastname: Zhao
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Xiao Xing
authors.initials: XX
authors.lastname: Lv
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Li
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Plastic and Burn Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, 710038, China.
authors.forename: Xue Yong
authors.initials: XY
authors.lastname: Li
mesh_terms: D000293:Adolescent; D000328:Adult; D000367:Age Factors; D001706:Biopsy; D018929:Cell Culture Techniques / Q000379:methods*; D002648:Child; D002675:Child, Preschool; D006801:Humans; D007223:Infant; D015603:Keratinocytes / Q000166:cytology*; D008875:Middle Aged; D012867:Skin / Q000166:cytology
keywords: CEA; CK19; cell behavior; culture time; keratinocyte