Windmolders S, Willems L, Daniëls A, Linsen L, Fanton Y, Hendrikx M, Koninckx R, Rummens J-L, Hensen K
Laboratory of Experimental Hematology, Jessa Hospital, 3500, Hasselt, Belgium; Faculty of Medicine and Life Sciences, Hasselt University, 3590, Diepenbeek, Belgium.
Cell Prolif. 2015 Apr;48(2):175-86. doi: 10.1111/cpr.12166. Epub 2015 Jan 29.
Cardiac atrial appendage stem cells (CASCs) have recently emerged as an attractive candidate for cardiac regeneration after myocardial infarction. As with other cardiac stem cells, CASCs have to be expanded ex vivo to obtain clinically relevant cell numbers. However, foetal calf serum (FCS), which is routinely used for cell culturing, is unsuitable for clinical purposes, and influence of long-term in vitro culture on CASC behaviour is unknown.
We examined effects on CASC biology of prolonged expansion, and evaluated a culture protocol suitable for human use.
In FCS-supplemented medium, CASCs could be kept in culture for 55.75 ± 3.63 days, before reaching senescence. Despite a small reduction in numbers of proliferating CASCs (1.37 ± 0.52% per passage) and signs of progressive telomere shortening (0.04 ± 0.02 kb per passage), their immunophenotype and myocardial differentiation potential remained unaffected during the entire culture period. The cells were successfully expanded in human platelet plasma supernatant, while maintaining their biological properties.
We successfully developed a protocol for long-term culture, to obtain clinically relevant CASC numbers, while retaining their cardiogenic potential. These insights in CASC biology and optimization of a humanized platelet-based culture method are an important step towards clinical application of CASCs for cardiac regenerative medicine.
心房附属器干细胞（CASC）最近已成为心肌梗死后心脏再生的一个有吸引力的候选细胞。与其他心脏干细胞一样，CASC必须在体外进行扩增，以获得临床上相关的细胞数量。然而，常规用于细胞培养的胎牛血清（FCS）不适合临床应用，且长期体外培养对CASC行为的影响尚不清楚。
我们研究了长期扩增对CASC生物学特性的影响，并评估了一种适合人类使用的培养方案。
在添加FCS的培养基中，CASC在达到衰老前可在培养中维持55.75±3.63天。尽管增殖的CASC数量略有减少（每传代1.37±0.52%），且有端粒逐渐缩短的迹象（每传代0.04±0.02 kb），但其免疫表型和心肌分化潜能在整个培养期间仍未受影响。这些细胞在人血小板血浆上清液中成功扩增，同时保持其生物学特性。
我们成功开发了一种长期培养方案，以获得临床上相关的CASC数量，同时保留其心脏生成潜能。这些关于CASC生物学特性的见解以及基于人血小板的培养方法的优化是CASC在心脏再生医学临床应用的重要一步。
