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Metabolic and proliferation evaluation of human adipose-derived mesenchymal stromal cells (ASC) in different culture medium volumes: standardization of static culture.
Biologicals ( IF 1.5 ) Pub Date : 2019-09-06 , DOI: 10.1016/j.biologicals.2019.08.006
Vinícius Augusto Simão 1 , Caetano Pedro Evangelista-Ribeiro 2 , Heloisa Brand 2 , Lougan Lagass-Pereira 3 , Laís Fernanda Marques 1 , Pedro Henrique Benites-Aoki 2 , Roseli Nunes da Silveira-Antunes 4 , Aldo Tonso 5 , João Tadeu Ribeiro-Paes 2
Affiliation  

Adipose-derived mesenchymal stromal/stem cells (ASC) have acquired a prominent role in tissue engineering and regenerative medicine. However, the standardization of basic culture procedures in this cellular type is still not well established according to the main qualitative cellular attributes. We evaluate the cell growth profile of human ASC in a different culture medium volumes and their nutritional composition utilizing static cultivation. Culture medium volumes (5, 10 and 15 mL/25 cm2) in T-flasks were evaluated by kinetic parameters and the metabolic composition was determined by biochemical analysis and Fourier transform infrared (FT-IR) absorption spectroscopy. 50% renewal of culture medium volume every 48 h was adopted. Immunophenotypic characterization and cell differentiation were performed. There was no difference (p > 0.05) in the kinetic parameters of cell proliferation between the culture medium volumes or in FT-IR composition. However, the concentrations of glucose, glutamine, lactate, and glutamate varied significantly during the cultivation process as a function of the medium volume. ASC presented specific antigens and differentiation potential of mesenchymal stromal/stem cells. It was concluded that the minimal culture medium volume (5 mL/25 cm2 in static culture) was sufficient to maintain the stability, potency, and growth of ASC, representing an economic and safe standardization for this cell culture process.



中文翻译:

不同培养基体积中人类脂肪间充质基质细胞(ASC)的代谢和增殖评估:静态培养的标准化。

脂肪来源的间充质基质/干细胞(ASC)在组织工程和再生医学中已发挥了重要作用。但是,根据主要的定性细胞属性,仍不能很好地建立这种细胞类型的基本培养程序的标准化。我们评估了人类ASC在不同培养基体积中的细胞生长概况及其利用静态培养的营养成分。培养基体积(5、10和15 mL / 25 cm 2通过动力学参数评估T瓶中的),并通过生化分析和傅里叶变换红外(FT-IR)吸收光谱法确定代谢成分。每48小时采用50%的培养基体积更新。进行了免疫表型鉴定和细胞分化。培养基体积之间或FT-IR组成方面,细胞增殖的动力学参数没有差异(p> 0.05)。但是,葡萄糖,谷氨酰胺,乳酸盐和谷氨酸的浓度在培养过程中随培养基体积的变化而显着变化。ASC呈现间充质基质/干细胞的特异性抗原和分化潜能。结论是最小培养基体积(5 mL / 25 cm 2 在静态培养中)足以维持ASC的稳定性,效力和生长,代表此细胞培养过程的经济和安全标准化。

更新日期:2019-09-06
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