细胞培养评估 4°C 下在等渗溶液和营养介质中储存的网切断层皮片的质量。
Quality evaluation of meshed split-thickness skin grafts stored at 4°C in isotonic solutions and nutrient media by cell cultures.
机构信息
Skin Laboratory, Burns Unit, Concord Hospital, Concord, NSW 2139, Australia.
出版信息
Burns. 2012 Sep;38(6):899-907. doi: 10.1016/j.burns.2012.02.002. Epub 2012 Mar 3.
OBJECTIVE
Excess split-skin autografts harvested and meshed during burn surgery are often stored at 4°C temporarily for later use. The quality of the stored skin is critical to clinical outcome and needs to be assured.
METHODS
Meshed split-thickness skin graft (mSSG) stored in saline, Hartmann's solution and two cell culture media, Dulbecco's Modified Eagle Medium (DMEM) and DMEM/Ham F12 (DMEM/F12, 3:1 mixture) were analyzed by trypan blue staining, cell culture and microbiological testing through a 28-day time course for cell viability and microbial contamination.
RESULTS
mSSG samples in all groups showed a progressive decrease of cell viability and colony forming efficiency through the time course of storage at 4°C. Cell culture media were better than saline and Hartmann's solution in maintaining the viability and growth capability of skin cells. The viability observed by trypan blue staining did not truly reflect the cell growth capacity after storage. mSSG in saline and Hartman's solution retained minimal keratinocyte growth potency after 7 days. mSSG in cell culture media had significant loss of keratinocyte colony growth potency after 7 days and minimal keratinocyte growth after 14 days. Dermal fibroblasts of all groups were less tolerant than keratinocytes to the storage. Microbial contaminations were common in mSSG harvested from burn surgery.
CONCLUSIONS
Culture media instead of saline or Hartman's solution should be used for temporary storage of mSSG at 4°C. The stored mSSS should be used within seven days to have sufficient viable number and cell growth efficiency. After then, the efficacy of stored mSSG as a source of living cells for wound closure could be full of uncertainty due to significant decrease of keratinocyte colony forming efficiency. Precaution should be taken during skin harvest and storage to minimize the risk of sample contamination. Inclusion of antimicrobial agents in storage solution and microbiological testing are advisable to ensure the quality and clinical outcome.
目的
在烧伤手术中采集和网切的多余的刃厚皮片通常在 4°C 下临时储存以备后用。储存皮肤的质量对临床结果至关重要，需要得到保证。
方法
将储存于生理盐水、哈特曼溶液和两种细胞培养基（DMEM 和 DMEM/F12，3:1 混合物）中的网切刃厚皮片（mSSG）进行台盼蓝染色、细胞培养和微生物检测，通过 28 天的时间过程分析细胞活力和微生物污染。
结果
在 4°C 下储存的 mSSG 样本在整个时间过程中表现出细胞活力和集落形成效率逐渐下降。细胞培养基比生理盐水和哈特曼溶液更能维持皮肤细胞的活力和生长能力。台盼蓝染色观察到的活力并不能真实反映储存后的细胞生长能力。在 7 天后，生理盐水和哈特曼溶液中的 mSSG 仅保留最小的角质形成细胞生长能力。在细胞培养基中的 mSSG 在 7 天后丧失了显著的角质形成细胞集落生长能力，在 14 天后几乎没有角质形成细胞生长。所有组的真皮成纤维细胞对储存的耐受性均低于角质形成细胞。从烧伤手术中采集的 mSSG 中普遍存在微生物污染。
结论
mSSG 在 4°C 下临时储存时应使用培养基而不是生理盐水或哈特曼溶液。储存的 mSSS 应在 7 天内使用，以保证有足够的活细胞数量和细胞生长效率。在此之后，由于角质形成细胞集落形成效率显著降低，储存的 mSSG 作为伤口闭合的活细胞来源的疗效可能充满不确定性。在皮肤采集和储存过程中应采取预防措施，以最大程度地降低样本污染的风险。在储存溶液中加入抗菌剂和进行微生物检测是确保质量和临床结果的明智之举。
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