CAJ | 학술논문

目的：判断深低温冷冻保存对于脐血单个核细胞和CD34+细胞集落形成能力及凋亡率的影响，从而选取能准确反映冷冻损伤的指标以及优化脐血冷冻保存的方式。方法：从脐血中分离出单个核细胞及CD34+细胞，对这两种形式的细胞进行深低温冷冻保存，保存30 d后复温，并分别进行冷冻前后的粒-巨噬细胞集落形成单位(colony forming unit-granulocyte/monocyte，CFU-GM)和髓系多向造血干细胞(colony forming unit-granulocyte，erythrocyte, monocyte and megakaryocyte，CFU-GEMM)的培养，比较冷冻前后的细胞集落产率，同时Annexin V-FITC-PI染色后流式细胞仪检测两种保存形式的细胞冷冻前后的细胞凋亡率。结果：冷冻保存对于以单个核细胞形式保存的细胞其集落形成能力和以CD34+细胞形式保存细胞的CFU-GM产率均无明显影响(P=0.31)，而对以CD34+细胞形式保存的细胞其CFU-GEMM的产率却有明显影响，冷冻后的CFU-GEMM的产率明显低于冷冻前(P<0.05)，且冷冻前即发现两种形式的细胞均有凋亡，冷冻后以单个核细胞形式保存的细胞其细胞凋亡率稍有所上升，但以CD34+细胞形式保存的细胞其细胞凋亡率更高。结论：冷冻前细胞即存在凋亡，冷冻对于以CD34+细胞形式冷冻保存的细胞中的较早期的祖细胞CFU-GEMM存在一定的损伤，其损伤可能主要为诱导细胞凋亡所致，而对于以单个核细胞形式保存的细胞的影响较小。
목적：판단심저온냉동보존대우제혈단개핵세포화CD34+세포집락형성능력급조망솔적영향，종이선취능준학반영냉동손상적지표이급우화제혈냉동보존적방식。방법：종제혈중분리출단개핵세포급CD34+세포，대저량충형식적세포진행심저온냉동보존，보존30 d후복온，병분별진행냉동전후적립-거서세포집락형성단위(colony forming unit-granulocyte/monocyte，CFU-GM)화수계다향조혈간세포(colony forming unit-granulocyte，erythrocyte, monocyte and megakaryocyte，CFU-GEMM)적배양，비교냉동전후적세포집락산솔，동시Annexin V-FITC-PI염색후류식세포의검측량충보존형식적세포냉동전후적세포조망솔。결과：냉동보존대우이단개핵세포형식보존적세포기집락형성능력화이CD34+세포형식보존세포적CFU-GM산솔균무명현영향(P=0.31)，이대이CD34+세포형식보존적세포기CFU-GEMM적산솔각유명현영향，냉동후적CFU-GEMM적산솔명현저우냉동전(P<0.05)，차냉동전즉발현량충형식적세포균유조망，냉동후이단개핵세포형식보존적세포기세포조망솔초유소상승，단이CD34+세포형식보존적세포기세포조망솔경고。결론：냉동전세포즉존재조망，냉동대우이CD34+세포형식냉동보존적세포중적교조기적조세포CFU-GEMM존재일정적손상，기손상가능주요위유도세포조망소치，이대우이단개핵세포형식보존적세포적영향교소。
Objective:To evaluate the effect of cryopreservation on clonogenic ability and apoptosis rate of mono-nuclear cells and CD34+cells in umbilical blood (UB), and to choose the index to present the freezing injury and optimize the cryopreservation of UB. Methods:hTe mono-nuclear cells (MNC) and CD34+cells were separated from UB and frozen.Atfer 30 days, they were thawed in warm water. Clonogenic capacity and clonogenic recovery before and atfer the cryopreservation was compared. We also used Annexin V-FITC-PI to investigate the apoptosis rate of the cells before and atfer the cryopreservation of these 2 types of cells. Results:hTe number of colony forming unit-granulocyte/monocyte (CFU-GMs) was not changed atfer freezing and thawing in both MNCs and CD34+cells, while the number of colony forming unit-granulocyte, erythrocyte, monocyte and megakaryocyte (CFU-GEMM) was obviously reduced after freezing in CD34+cells. The 2 types of cryopreserved cells had certain degree of apoptosis before the cryopreservation. MNC-type cryopreservation increased the cells apoptosis a little, while CD34+-type cryopreservation increased more. Conclusion:hTe cells have certain degree of apoptosis before the cryopreservation. hTe freezing and thawing procedure does affect the early stage progenitor cells-CFU-GEMM in the CD34+-type cryopreserved cells in UB. hTe damage may be induced by the cell apoptosis.