Wang H J, Chen T M, Cheng L F, Cheng T Y, Tung Y M
Department of Surgery, Tri-service General Hospital, Taipei, Taiwan, Republic of China.
Burns. 1995 Nov;21(7):503-6. doi: 10.1016/0305-4179(95)00030-f.
Extensive burn wound coverage remains a major problem in acute and reconstructive stages due to shortage of autologous skin donor sites. Many reports have described successful coverage of extensive burn wounds by using cultured keratinocytes sheet graft alone or overlaying a glycosaminoglycan substrate, allodermis or xenodermis. In our experience, the use of Boyce and Ham's serum-free medium made the preparation of keratinocyte cultures simple and the cell growth was prompt. In this study, one of the supplements in Boyce's culture medium, the bovine pituitary extract (BPE at concentration of 140 micrograms/ml), was replaced by different concentrations of fresh frozen porcine pituitary extract (PPE). The protein concentration of the original preparation of PPE was also 140 micrograms/ml but in which 375 +/- 21 pg/ml (1/10 dilution) of platelet-derived growth factor (PDGF) was detected by immunoassay (PDGF ELISA). Our results proved that PPE is an ideal supplement for the in vitro growth of keratinocytes, and when its protein concentration is 350 micrograms/ml will induce a significant keratinocyte growth by day 9 when compared to the regular BPE and also to the other concentrations of PPE (P < 0.05).
由于自体皮肤供区不足，大面积烧伤创面的覆盖在急性期和修复期仍然是一个主要问题。许多报告描述了单独使用培养的角质形成细胞片移植或覆盖糖胺聚糖底物、同种异体真皮或异种真皮成功覆盖大面积烧伤创面的情况。根据我们的经验，使用博伊斯和哈姆的无血清培养基使角质形成细胞培养的制备变得简单，细胞生长迅速。在本研究中，博伊斯培养基中的一种补充剂，即浓度为140微克/毫升的牛垂体提取物（BPE），被不同浓度的新鲜冷冻猪垂体提取物（PPE）所取代。PPE原液的蛋白质浓度也是140微克/毫升，但通过免疫测定（PDGF ELISA）检测到其中含有375±21皮克/毫升（1/10稀释）的血小板衍生生长因子（PDGF）。我们的结果证明，PPE是角质形成细胞体外生长的理想补充剂，当其蛋白质浓度为350微克/毫升时，与常规BPE以及其他浓度的PPE相比，在第9天时将诱导角质形成细胞显著生长（P<0.05）。
