CAJ | 학술논문

背景:人脐带间充质干细胞作为新的种子细胞来源,如何迅速提高其增殖以满足临床所需的数量和质量,是迫切需要解决的现实问题.目的:探讨不同的灌注启动时间、灌注频率和灌注体积对人脐带间充质干细胞扩增和代谢的影响,建立高密度培养的半连续灌注培养工艺.方法:培养传代人脐带间充质干细胞,以批培养方式为参照,半连续灌注培养分成3组.①灌注启动时间组:批培养24 h 和48 h,分别开始换液,每2 d 更换75%.②灌注频率组:批培养48 h 开始换液,频率分别为每1,2,3 d,换液体积比为75%.③灌注体积组:批培养48 h 开始换液,频率为每2 d,换液体积比分别为25%,50%,75%,100%.结果与结论:相对于其他培养条件,灌注启动时间为48 h、频率为每天、体积比为75%的静态培养条件下,稀释率较大,细胞密度较大,葡萄糖比消耗速率和乳酸比生成速率较高,谷氨酰胺比消耗速率较低,氨比生成速率较高,乳酸对葡萄糖的代谢系数较低,谷氨酰胺的代谢系数较高.这表明稀释率大,不仅葡萄糖和谷氨酰胺充足,而且细胞所需的微量元素和未知的因子也充足,满足了细胞生长的要求;同时排除乳酸和氨等对细胞的影响,去除条件培养基中某些因子对细胞的交互影响,提高了细胞的增殖.
배경:인제대간충질간세포작위신적충자세포래원,여하신속제고기증식이만족림상소수적수량화질량,시박절수요해결적현실문제.목적:탐토불동적관주계동시간、관주빈솔화관주체적대인제대간충질간세포확증화대사적영향,건립고밀도배양적반련속관주배양공예.방법:배양전대인제대간충질간세포,이비배양방식위삼조,반련속관주배양분성3조.①관주계동시간조:비배양24 h 화48 h,분별개시환액,매2 d 경환75%.②관주빈솔조:비배양48 h 개시환액,빈솔분별위매1,2,3 d,환액체적비위75%.③관주체적조:비배양48 h 개시환액,빈솔위매2 d,환액체적비분별위25%,50%,75%,100%.결과여결론:상대우기타배양조건,관주계동시간위48 h、빈솔위매천、체적비위75%적정태배양조건하,희석솔교대,세포밀도교대,포도당비소모속솔화유산비생성속솔교고,곡안선알비소모속솔교저,안비생성속솔교고,유산대포도당적대사계수교저,곡안선알적대사계수교고.저표명희석솔대,불부포도당화곡안선알충족,이차세포소수적미량원소화미지적인자야충족,만족료세포생장적요구;동시배제유산화안등대세포적영향,거제조건배양기중모사인자대세포적교호영향,제고료세포적증식.
@@@@BACKGROUND: How to quickly improve the proliferation of human umbilical cord-derived mesenchymal stem cells as a seed cellsource is an urgent problem to meet the clinical requirement for the quantity and quality. OBJECTIVE: To explore the effects of different initial time, frequencies and volumes of perfusion on the proliferation and metabolism of human umbilical cord-derived mesenchymal stem cells and to establish a semi-continuous perfusion culture technique with high cel density. METHODS: Human umbilical cord-derived mesenchymal stem cells were cultured and passaged, the batch cultured cells were used as control. The semi-continuous perfusion cultured cells were divided into three groups. In the initial time group: the cells were batch cultured for 24 and 48 hours, respectively, and then the media was changed for 75% every 2 days. In the perfusion frequency group: the cells were batch cultured for 48 hours, after that, the media was changed every 1, 2 and 3 days, respectively, and 75% media was changed. In the perfusion volume group: the media was changed every 2 days after batch cultured for 48 hours, 25%, 50%, 75% and 100%media was changed. RESULTS AND CONCLUSION: The relatively higher values of dilution rate and cel density were acquired with the suitable monolayer culture of perfusion initial time at 48 hours of batch culture, and 75% (volume ratio) media exchanged every day. The higher values of glucose consumption rate, lactate formation rate and ammonia formation rate, and lower value of glutamine consumption rate were attained with higher metabolic coefficient of lactate/glucose and lower metabolic coefficient of ammonia/glutamine. It showed that with higher medium dilution, nutrients including glucose, glutamine, trace elements and other unknown factors were sufficiently supplied to meet the requirement of cel growth, and the negative effects of lactate and ammonium and the interaction effect of some factors in the culture media on the cells were relieved, leading to the enhanced proliferation of cells.