联合使用干细胞因子、FLT3配基与白介素2,7,15体外扩增人脐血来源CIK/NK细胞

Expansion of CIK/NK Cells from Cord Blood by Using Different Combinations of Stem Cell Factor,FLT3 Ligand and Interleukin 2,7,15 in vitro

通过干细胞因子 (SCF)、FLT3配基 (FL)联合白介素 (IL) 2 ,7,15等的不同组合体系体外扩增人脐血来源的CIK NK细胞 (CB CIK NK) ,探讨不同培养方式对CB CIK NK细胞诱导、扩增产率的影响。按诱导扩增体系的不同分 3组 :A组 (SCF +IL2 +IL7+IL15 ) ,B组 (SCF +FL +IL2 +IL7+IL15 )和C组 (IL2 +IL7+IL15 ,对照组 ) ,培养 2 1天 ,检测CD3+ CD5 6 + CIK细胞、CD3- CD5 6 + NK细胞比例和扩增倍数。结果表明 :经 2 1天培养 ,A ,B及C组CIK细胞比例分别为 (19.84± 2 .93) %、(2 6 .2 0± 4 .0 5 ) %及 (2 4 .0 3± 4 .99) % ;而NK细胞的比例A、B及C组比例分别为 (4 9.6 0± 1.4 0 ) %、(5 1.16± 6 .4 5 ) %及 (30 .85± 8.12 ) %。含SCF +FL的B组扩增效率最高 ,其中CIK细胞的扩增倍数由对照组 (C组 )的 (5 75 81± 2 2 1.72 )倍增加至 (796 .0 9± 2 78.4 7)倍 (最高为 1313倍 ) ;NK细胞由对照组 (C组 )的 (30 .39± 14 .4 7)倍增加至 (6 5 .85± 30 .83)倍 (最高为 12 1.0 6倍 )。结论 :通过合适的细胞因子组合可以同时扩增CB来源的CIK NK细胞 ;联合诱导、扩增CB CIK NK细胞以采用含SCF +FL +IL2 +IL7+IL15的培养体系为佳。

To explore the efficacy of the different combinations of stem cell factor (SCF),FLT3 ligand(FL)and interleukin (IL)2,7 ,15 on the induction and expansion of cord blood (CB) derived CIK/ NK cells in vitro,according to the different combinations of cytokines,combinations of cytokines were divided into 3 groups: group A (SCF+IL2+IL7+IL15),group B (SCF+FL+IL2+IL7+IL15) and group C (IL2+IL7+IL15 as control group). At 21 days of culture,the proportion and the expansion rate of CD3+CD56+ CIK cells and CD3-CD56+ NK cells were measured. The results showed that at 21 days in culture,the percentages of CD3+CD56+ CIK cells derived from CB-MNC in group A,B and C were (19.84±2.93)%,(26.20±4.05)%,(24.03±4.99%) and that of NK cells were (49.60±1.40)%,(51.16±6.45)% and (30.85±8 12)%,respectively. The proliferation of CD3+CD56+ CIK cells and CD3-CD56+ NK cells was the most effective in group B (SCF+FL+IL2+IL7+IL15). Expansion multiple of CIK cell number increased from 575.81±221.72 (control) to 796.09±278.47 with peak value of 1 313 ,and that of NK cells increased from 30.39±14.47 (control) to 65.85±30.83 with peak value of 121.06. In conclusion,a proper cytokines (SCF+FL+IL2+IL7+IL15) culture system can effectively induce and expand CB CD3+CD56+ CIK cells and CD3-CD56+ NK cells simultaneously.