Abstract:
The absence of a native extracellular matrix and the use of xenogeneic sera are often associated with rapid tenocyte function losses during in vitro culture. Herein, we assessed the influence of different sera (equine serum and foetal bovine serum) on equine tenocyte morphology, viability, metabolic activity, proliferation and protein synthesis as a function of tissue-specific extracellular matrix deposition (induced via macromolecular crowding), aging (passages 3, 6, 9) and time in culture (days 3, 5, 7). In comparison to cells at passage 3, at day 3, in foetal bovine serum and without macromolecular crowding (traditional equine tenocyte culture), the highest number of significantly decreased readouts were observed for cells in foetal bovine serum, at passage 3, at day 5 and day 7 and without macromolecular crowding. Again, in comparison to traditional equine tenocyte culture, the highest number of significantly increased readouts were observed for cells in equine serum, at passage 3 and passage 6, at day 7 and with macromolecular crowding. Our data advocate the use of an allogeneic serum and tissue-specific extracellular matrix for effective expansion of equine tenocytes.
摘要:
天然细胞外基质的缺乏和异种血清的使用通常与体外培养过程中肌腱细胞功能的快速丧失有关。在这里,我们评估了不同血清(马血清和胎牛血清)对马肌腱细胞形态的影响,生存能力,代谢活动,增殖和蛋白质合成作为组织特异性细胞外基质沉积的功能(通过大分子拥挤诱导),老化(第3、6、9代)和培养时间(第3、5、7天)。与第3代的细胞相比,在第3天,在胎牛血清中并且没有大分子拥挤(传统的马肌腱细胞培养),胎牛血清中观察到的细胞读数显着降低的最高数量,在第3天,第5天和第7天,没有大分子拥挤。再一次,与传统的马肌腱细胞培养相比,在马血清中观察到的细胞读数显着增加的最高数量,在第3次和第6次,第7天,大分子拥挤。我们的数据主张使用同种异体血清和组织特异性细胞外基质有效扩增马肌腱细胞。