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人血小板裂解物与胎牛血清比较——研究用于临床的人脂肪组织来源干细胞的增殖率、染色体稳定性和血管生成潜力。

Pooled human platelet lysate versus fetal bovine serum-investigating the proliferation rate, chromosome stability and angiogenic potential of human adipose tissue-derived stem cells intended for clinical use.

机构信息

Department of Plastic Surgery, Breast Surgery & Burns 2102, Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark.

出版信息

Cytotherapy. 2013 Sep;15(9):1086-97. doi: 10.1016/j.jcyt.2013.01.217. Epub 2013 Apr 17.

Abstract

BACKGROUND AIMS

Because of an increasing focus on the use of adipose-derived stem cells (ASCs) in clinical trials, the culture conditions for these cells are being optimized. We compared the proliferation rates and chromosomal stability of ASCs that had been cultured in Dulbecco's modified Eagle's Medium (DMEM) supplemented with either pooled human platelet lysate (pHPL) or clinical-grade fetal bovine serum (FBS) (DMEM(pHPL) versus DMEM(FBS)).

METHODS

ASCs from four healthy donors were cultured in either DMEM(pHPL) or DMEM(FBS), and the population doubling time (PDT) was calculated. ASCs from two of the donors were expanded in DMEM(pHPL) or DMEM(FBS) and cultured for the final week before harvesting with or without the addition of vascular endothelial growth factor. We assessed the chromosomal stability (through the use of array comparative genomic hybridization), the expression of ASC and endothelial surface markers and the differentiation and angiogenic potential of these cells.

RESULTS

The ASCs that were cultured in pHPL exhibited a significantly shorter PDT of 29.6 h (95% confidence interval, 22.3-41.9 h) compared with those cultured in FBS, for which the PDT was 123.9 h (95% confidence interval, 95.6-176.2 h). Comparative genomic hybridization analyses revealed no chromosomal aberrations. Cell differentiation, capillary structure formation and cell-surface marker expression were generally unaffected by the type of medium supplement that was used or by the addition of vascular endothelial growth factor.

CONCLUSIONS

We observed that the use of pHPL as a growth supplement for ASCs facilitated a significantly higher proliferation rate compared with FBS without compromising genomic stability or differentiation capacity.

摘要

背景目的

由于越来越关注脂肪来源干细胞(ASCs)在临床试验中的应用,这些细胞的培养条件正在得到优化。我们比较了在补充有混合人血小板裂解物(pHPL)或临床级胎牛血清(FBS)的 DMEM 中培养的 ASC 的增殖率和染色体稳定性(DMEM(pHPL)与 DMEM(FBS))。

方法

从四位健康供体中培养 ASC,分别在 DMEM(pHPL)或 DMEM(FBS)中培养,并计算群体倍增时间(PDT)。两位供体的 ASC 在 DMEM(pHPL)或 DMEM(FBS)中扩增,并在收获前的最后一周添加或不添加血管内皮生长因子进行培养。我们评估了染色体稳定性(通过使用阵列比较基因组杂交)、ASC 和内皮表面标志物的表达以及这些细胞的分化和血管生成潜力。

结果

在 pHPL 中培养的 ASC 的 PDT 显著缩短至 29.6 h(95%置信区间,22.3-41.9 h),而在 FBS 中培养的 PDT 为 123.9 h(95%置信区间,95.6-176.2 h)。比较基因组杂交分析显示没有染色体异常。细胞分化、毛细血管结构形成和细胞表面标志物表达通常不受培养基补充物类型或添加血管内皮生长因子的影响。

结论

我们观察到,与 FBS 相比,使用 pHPL 作为 ASC 的生长补充剂可显著提高增殖率,而不会影响基因组稳定性或分化能力。

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