CAJ | 학술논문

目的了解烧伤水疱液对人骨髓间充质于细胞(MSC)体外培养及表型转化的影响.方法体外分离、培养、扩增、鉴定人骨髓MSC,收集烧伤患者伤后12、24、48 h水疱液,分别加入MSC培养液中.分为水疱液1组:MSC培养液中加入伤后12 h水疱液;水疱液2组:MSC培养液中加入伤后24 h水疱液;水疱液3组:MSC培养液中加入伤后48 h水疱液;对照组:常规培养.各组所加水疱液体积分数均为20%.观察水疱液微生物生长情况及各组MSC生长情况.流式细胞仪检测培养8 d时各组MSC中CD44、细胞角蛋白7(CK7)阳性表达率.结果 15个水疱液标本细菌、真菌培养均为阴性;各组MSC细胞形态无明显变化,但各水疱液组细胞数量均少于对照组,水疱液3组细胞数量下降最明显.水疱液1、2、3组CD44阳性表达率分别为(83.0±3.1)%、(77.2±2.9)%、(65.1±2.3)%,均低于对照组[(89.5±3.2)%,P<0.01];水疱液1、2、3组CK7阳性表达率分别为(24.06±0.11)%、(16.41±0.09)%、(4.48±0.07)%,明显高于对照组[(3.87±0.04)%,P<0.01].结论烧伤创面水疱液对MSC体外培养有明显的抑制作用,并具有一定的促进MSC表型转化作用.
목적 료해소상수포액대인골수간충질우세포(MSC)체외배양급표형전화적영향.방법 체외분리、배양、확증、감정인골수MSC,수집소상환자상후12、24、48 h수포액,분별가입MSC배양액중.분위수포액1조:MSC배양액중가입상후12 h수포액;수포액2조:MSC배양액중가입상후24 h수포액;수포액3조:MSC배양액중가입상후48 h수포액;대조조:상규배양.각조소가수포액체적분수균위20%.관찰수포액미생물생장정황급각조MSC생장정황.류식세포의검측배양8 d시각조MSC중CD44、세포각단백7(CK7)양성표체솔.결과 15개수포액표본세균、진균배양균위음성;각조MSC세포형태무명현변화,단각수포액조세포수량균소우대조조,수포액3조세포수량하강최명현.수포액1、2、3조CD44양성표체솔분별위(83.0±3.1)%、(77.2±2.9)%、(65.1±2.3)%,균저우대조조[(89.5±3.2)%,P<0.01];수포액1、2、3조CK7양성표체솔분별위(24.06±0.11)%、(16.41±0.09)%、(4.48±0.07)%,명현고우대조조[(3.87±0.04)%,P<0.01].결론 소상창면수포액대MSC체외배양유명현적억제작용,병구유일정적촉진MSC표형전화작용.
Objective To study the effect of blister fluid obtained from burn patient on human MSCs in vitro and its phenotypic modulation in culture. Methods Blister fluid from burn patients was col-lected at 12, 24, 48 post burn hour (PBH). The human MSCs were isolated, cultured, amplified and iden-tified in vitro, then were divided into A (culture with 20% blister fluid collected at 12 PBH) , B (culture with 20% blister fluid collected at 24 PBH) , C (culture with 20% blister fluid collected at 48 PBH), N (with ordinary culture medium) groups. The growth of MSCs and micro-organisms in blister fluid were ob-served. Positive expression rates of CD44 and CK7 were detected by flow cytometry after culture for 8 days. Results Bacterial and fungal growths were absent in 15 blister fluid samples. There was no obvious change in MSC morphology in each group. Compared with that of N group, the number of MSCs in A, B, C groups was decreased, especially in C group. CD44 positive expression rate in A, B, C groups was ( 83.0± 3.1)%, (77.2±2.9)% and (65.1±2.3)%, respectively,which was obviously lower than that in N group [(89.5±3.2)%, P <0.01]. CK7 positive expression rate in A, B, C groups was (24.06± 0.11) % , (16.41±0.09) % and (4.48±0.07) % , respectively, which was obviously higher than that in N group [(3.87±0.04)% , P<0.01 ]. Conclusions Burn blister fluid can obviously inhibit the growth of human MSC cultured in vitro, and may promote modulation of its phenotype to certain extent.