Compton C C, Warland G, Kratz G
Department of Pathology, Massachusetts General Hospital, Boston 02114, USA.
J Burn Care Rehabil. 1998 Jul-Aug;19(4):330-6. doi: 10.1097/00004630-199807000-00011.
Patchy hypopigmentation often occurs unpredictably in the skin regenerated from cultured epidermal autografts, especially when that skin is grown from frozen cells, serially passaged, or both. The impact of serial subcultivation and cryopreservation on melanocyte viability in the cultured epidermal autograft culture system was investigated. Serial subcultivation of human keratinocytes through as many as eight passages was performed, and melanocyte densities in confluent cultures at each passage were determined after specific labeling of melanocytes. The experimental cells were frozen before cultivation and between passages to determine the effect of standard cryopreservation on melanocyte survival. Freshly passaged cells that had not been frozen served as controls. Melanocytes were gradually depleted during fresh passage of epidermal cells but persisted through as many as seven passages. Freezing before or after the first passage or between subsequent passages resulted in a complete loss of melanocytes by the third or fourth passage. The findings suggest that cryopreservation should be avoided during cultured epidermal autograft production to optimize melanocyte survival and minimize pigmentation abnormalities that may occur after grafting.
片状色素减退在培养的自体表皮移植物再生的皮肤中经常不可预测地出现，尤其是当该皮肤由冷冻细胞生长、连续传代或两者兼而有之时。研究了连续传代培养和冷冻保存对培养的自体表皮移植物培养系统中黑素细胞活力的影响。对人角质形成细胞进行多达八代的连续传代培养，并在对黑素细胞进行特异性标记后，测定每一代汇合培养物中的黑素细胞密度。在培养前和传代之间对实验细胞进行冷冻，以确定标准冷冻保存对黑素细胞存活的影响。未冷冻的新鲜传代细胞作为对照。黑素细胞在表皮细胞新鲜传代过程中逐渐减少，但可维持多达七代。在第一代传代之前或之后或在后续传代之间进行冷冻，会导致在第三代或第四代时黑素细胞完全丧失。研究结果表明，在培养的自体表皮移植物生产过程中应避免冷冻保存，以优化黑素细胞存活，并将移植后可能出现的色素沉着异常降至最低。
