CAJ | 학술논문

目的：观察补肾中药血清联合BMP?2对人脐血间充质干细胞（ UCB?MSCs）增殖及形态学的影响，并探讨其最佳时效。方法倒置显微镜观察原代、传代培养及干预后UCB?MSCs的形态学变化；取第3代UCB?MSCs，随机分为8组，给予不同浓度补肾中药血清及BMP?2条件培养基培养，分别于诱导后1 d、3 d、5 d采用MTT法检测各组细胞增殖情况。结果原代培养UCB?MSCs随时间延长，数量逐渐增多且逐渐变长，2周时多呈长梭形、旋涡状生长，传代培养3 d即达到70％～80％融合，以梭形为主，符合干细胞的生长特征；干预后第3、5 d，5％、10％中药＋BMP?2组和BMP?2组均能够促进UCB?MSCs增值（ P＜0?05），且5％中药＋ BMP?2组优于BMP?2组（P＝0?041），增殖高峰出现在干预后第3 d；经条件培养3 d后，各组细胞均可见由长梭形变成立方形或多角形，尤以5％中药联合BMP?2组及BMP?2组较为明显。结论补肾中药血清联合BMP?2可协同促进体外培养UCB?MSCs增殖，增殖效应与干预时间和中药血清浓度有一定相关性。
목적：관찰보신중약혈청연합BMP?2대인제혈간충질간세포（ UCB?MSCs）증식급형태학적영향，병탐토기최가시효。방법도치현미경관찰원대、전대배양급간예후UCB?MSCs적형태학변화；취제3대UCB?MSCs，수궤분위8조，급여불동농도보신중약혈청급BMP?2조건배양기배양，분별우유도후1 d、3 d、5 d채용MTT법검측각조세포증식정황。결과원대배양UCB?MSCs수시간연장，수량축점증다차축점변장，2주시다정장사형、선와상생장，전대배양3 d즉체도70％～80％융합，이사형위주，부합간세포적생장특정；간예후제3、5 d，5％、10％중약＋BMP?2조화BMP?2조균능구촉진UCB?MSCs증치（ P＜0?05），차5％중약＋ BMP?2조우우BMP?2조（P＝0?041），증식고봉출현재간예후제3 d；경조건배양3 d후，각조세포균가견유장사형변성립방형혹다각형，우이5％중약연합BMP?2조급BMP?2조교위명현。결론보신중약혈청연합BMP?2가협동촉진체외배양UCB?MSCs증식，증식효응여간예시간화중약혈청농도유일정상관성。
Objective To observe the effect of kidney?tonifying Chinese herb containing serum combined with BMP?2 on the morphology and proliferation of human umbilical cord blood mesenchymal stem cells ( UCB?MSCs) . Methods The morphology change of primary, passaged, and intervened UCB?MSCs was observed using an inverted microscope. The third passage of UCB?MSCs was randomly divided into 8 groups culturing in different concentrations of kidney?tonifying Chinese herb containing serum and BMP?2. The cell proliferation was detected using MTT method at 1?, 3?, and 5?day after the intervention. Results The number of primary UCB?MSCs increased and the shape enlarged gradually along with the culturing time. After 2 weeks, most of the UCB?MSCs were characterized by spiral shape growth. In 3 days after passage, 70% -80% of the cells fused in the culture, which met the growth characteristics of the stem cells. The cell proliferation in 5% and 10% combining groups and BMP?2 group increased after 3?and 5?day intervention (P<0?05). The effect in 5% combining group was superior to that in BMP?2 group (P=0?041). The proliferation peak appeared on the third day after the intervention. The UCB?MSCs became cubical or polygonal shape in 3 days after the conditioned culture, especially in 5% Chinese herb containing serum combined with BMP?2 group and BMP?2 group. Conclusion Kidney?tonifying Chinese herb containing serum combined with BMP?2 can synergistically promote the proliferation of UCB?MSCs. The stimulation is time and concentration dependent.