中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2008年
10期
679-683
,共5页
胎血%间充质干细胞%细胞分化%胰岛素%糖尿病
胎血%間充質榦細胞%細胞分化%胰島素%糖尿病
태혈%간충질간세포%세포분화%이도소%당뇨병
Umbilical cord blood%Mesenchymal stem cells%Cell differentiation%Insulin%Diabetes mellitus
目的 探讨脐血间充质干细胞(MSC)向胰岛素分泌细胞分化的潜能及其分化诱导条件.方法 分离脐血有核细胞,将其置于MesencultTM培养基中进行培养,并利用贴壁法进行纯化、扩增.扩增后的脐血MSC用表皮生长因子(EGF)、β-巯基乙醇和高糖进行诱导.采用RT-PCR、胰岛素免疫细胞化学染色方法对诱导后的细胞进行鉴定,定量检测胰岛素分泌水平及其对葡萄糖刺激的反应性;移植到链脲佐菌素诱导的糖尿病小鼠体内,观测其在体内的降糖作用.结果 诱导后细胞形态发生明显变化,细胞变圆且聚集成团;RT-PCR分析显示,细胞表达部分胰岛相关基因;细胞的胰岛素免疫细胞化学染色为阳性;而且细胞能分泌少量[(0.37±0.06)mU/L]胰岛素,并对糖刺激具有反应性,刺激指数为1.76;诱导后的细胞移植能降低糖尿病小鼠的血糖.结论 脐血MSC具有向胰岛素分泌细胞分化的潜能.
目的 探討臍血間充質榦細胞(MSC)嚮胰島素分泌細胞分化的潛能及其分化誘導條件.方法 分離臍血有覈細胞,將其置于MesencultTM培養基中進行培養,併利用貼壁法進行純化、擴增.擴增後的臍血MSC用錶皮生長因子(EGF)、β-巰基乙醇和高糖進行誘導.採用RT-PCR、胰島素免疫細胞化學染色方法對誘導後的細胞進行鑒定,定量檢測胰島素分泌水平及其對葡萄糖刺激的反應性;移植到鏈脲佐菌素誘導的糖尿病小鼠體內,觀測其在體內的降糖作用.結果 誘導後細胞形態髮生明顯變化,細胞變圓且聚集成糰;RT-PCR分析顯示,細胞錶達部分胰島相關基因;細胞的胰島素免疫細胞化學染色為暘性;而且細胞能分泌少量[(0.37±0.06)mU/L]胰島素,併對糖刺激具有反應性,刺激指數為1.76;誘導後的細胞移植能降低糖尿病小鼠的血糖.結論 臍血MSC具有嚮胰島素分泌細胞分化的潛能.
목적 탐토제혈간충질간세포(MSC)향이도소분비세포분화적잠능급기분화유도조건.방법 분리제혈유핵세포,장기치우MesencultTM배양기중진행배양,병이용첩벽법진행순화、확증.확증후적제혈MSC용표피생장인자(EGF)、β-구기을순화고당진행유도.채용RT-PCR、이도소면역세포화학염색방법대유도후적세포진행감정,정량검측이도소분비수평급기대포도당자격적반응성;이식도련뇨좌균소유도적당뇨병소서체내,관측기재체내적강당작용.결과 유도후세포형태발생명현변화,세포변원차취집성단;RT-PCR분석현시,세포표체부분이도상관기인;세포적이도소면역세포화학염색위양성;이차세포능분비소량[(0.37±0.06)mU/L]이도소,병대당자격구유반응성,자격지수위1.76;유도후적세포이식능강저당뇨병소서적혈당.결론 제혈MSC구유향이도소분비세포분화적잠능.
Objective To investigate the differential potential of mesenchymal stem cells(MSCs) derived from human umbilical cord blood (hUCB) into insulin-secreting cells and its inducing condition. Methods UCB nucleated cells (NCs) were isolated and cultured in MesencuhTM media. The obtained UCB MSC were purified by adherence method and expanded. Then they were induced with epidermal growth factor (EGF), B-mercaptoethanol and high concentration of glucose. The induced cells were identified by RT-PCR. Intracellular insulin was examined by immunocytochemistry. The quantity of insulin secretion and glucosesimulated insulin release were examined by chemiluminescence immtmoassay. The induced cells were also transplanted into renal subcapsular space of STZ-induced hyperglycemic mice to observe the in vivo lowering effect on hyperglycemia. Results The induced cells morphologically became round and were gathering into a mass. The expression of some genes related to pancreatic islet was found by RT-PCR. Chemiluminescence immunoassay showed insulin positivity and the cells secreted a low concentration of insulin [(0.37±0.06) mU/L]. The induced cells responded to high glucose challenge with a stimulation index of 1.76. Mter those cells grafted into renal subcapsularly there was an in vivo lowering effect on blood glucose level on STZ hyperglycemic mice. Conclusion MSCs from UCB can differentiated into insulin secreting cells.
