欢迎来到《四川大学学报(医学版)》

脂肪组织分泌物中蛋白浓度对诱导脂肪干细胞成脂的影响

Influence of Fetal Bovine Serum Protein Concentration in Adipose Tissue Extract on Adipogenic Differentiation of Adipose Derived Stem Cells

  • 摘要: 目的 观察胎牛血清(FBS)对体外培养获取的脂肪组织分泌物(ATE)诱导大鼠脂肪干细胞(ADSCs)成脂能力有无影响,并探讨ATE中不同蛋白浓度对诱导大鼠ADSCs成脂、细胞增殖和迁移能力的影响。 方法 培养大鼠ADSCs并传代,对ADSCs成骨和成神经诱导,采用茜素红染色和神经丝蛋白(NF)免疫荧光检测鉴定。含有FBS和无FBS的培养基对脂肪组织块进行培养并收集ATE,对第4代ADSCs进行诱导,第7 d时油红染色鉴定并计算成脂率;收集无FBS培养基获取的ATE,分为不同蛋白质量浓度组(100 μg/mL、250 μg/mL、500 μg/mL组),对第4代ADSCs进行成脂诱导,观察各组成脂率;并观察不同蛋白质量浓度组对ADSCs细胞增殖和迁移的影响。 结果 培养的ADSCs成骨和成神经诱导后茜素红染色和NF免疫荧光染色阳性;获取ATE时添加或不添加FBS,在第3 d时均能诱导ADSCs成脂,第7 d时的成脂率差异无统计学意义;ATE 500 μg/mL组较100 μg/mL、250 μg/mL组成脂率高(P均<0.05),细胞培养2 d后,3个不同蛋白质量浓度组均抑制细胞增殖,不同蛋白质量浓度间细胞抑制率差异无统计学意义(P均>0.05);ATE中不同蛋白质量浓度对细胞的迁移无影响。 结论 无FBS的培养基获取的ATE,短期内对ADSCs成脂能力没有影响;ATE蛋白浓度与ADSCs成脂率有关。

     

    Abstract: Objective To observe whether fetal bovine serum (FBS) will affect the adipogenic ability of adipose derived stem cells (ADSCs) induced by adipose tissue extract (ATE), and to explore the effects of different FBS protein concentrations in ATE on adipogenic ability, cell proliferation and migration of ADSCs. Methods Rat ADSCs were cultured, passaged, and then subjected to osteogenic and neural induction. The adipose explants were cultured in culture medium with or without FBS, then ATE was collected to induce passage four (P4)-ADSCs, which were subsequently detected with oil red staining on the 7th day and the adipogenic ratio was calculated. Different concentrations of ATE without FBS (100 μg/mL, 250 μg/mL and 500 μg/mL) were used to induce P4-ADSCs before the adipogenic events, and the adipogenic ratio of each concentration group was observed. The effect of different protein concentrations on ADSCs proliferation and migration was also observed. Results Alizarin red staining and NF immunofluorescence staining were positive after the osteogenic and neural induction of cultured ADSCs. Either ATE with or without FBS was able to induce adipogenic differentiation of ADSCs on the 3rd day and there was no significant differences of adipogenic differentiation between ATE with FBS and without FBS on the 7th day. The adipogenic ratio of 500 μg/mL group was higher than that of 100 μg/mL group and 250 μg/mL group (P<0.05). After two days of induction, all the three different protein concentrations could inhibit cell proliferation, and different protein concentrations in ATE had no effect on the migration of ADSCs. Conclusion ATE obtained from culture medium without FBS has no effect on adipogenic capacity of ADSCs in a short period of time. The adipogenic ratio of ADSCs is associated with protein concentration in ATE.

     

/

返回文章
返回