Carter J, Karmiol S, Nagy M, McElreath R, Calloway C, Motley A, Neill A, Jang H J, Posselt A, Stock P
Transplantation Research Lab, University of California-San Francisco, San Francisco, CA 94143, USA.
Transplant Proc. 2005 Oct;37(8):3446-9. doi: 10.1016/j.transproceed.2005.09.073.
Human islet transplant protocols frequently incorporate a brief period of islet culture before transplantation. The optimal medium for pretransplant islet culture is unknown.
We compared four serum-free media formulated for human islets: Miami (MM1), Memphis (M-SFM), Edmonton (EDM), and hCell OCZEM-SF/AF (hCell). Islets isolated from a single human pancreas with purity >80% were cultured in 2500-islet-equivalent (IE) fractions using the media listed. After 7 days, each 2500-IE fraction was grafted under the kidney capsule of a streptozocin-diabetic rag1 mouse (n = 4 per group). Mice were evaluated with serum glucose monitoring, stimulated C-peptide release, and glucose tolerance tests. Islet fractions transplanted immediately after isolation (n = 4 mice) served as controls. In vitro islet function was assessed on days 0 and 3 and included insulin release (after static glucose stimulation), total cellular C-peptide content, cell count, and viability.
Glucose control was improved in all cohorts of mice after transplant, but only islet grafts cultured in MM1 were statistically indistinguishable from fresh islets. MM1- and hCell-cultured islet grafts showed improved glucose tolerance compared with fresh islets; C-peptide release was similar among the four cohorts. In vitro, only islets cultured in MM1 had similar stimulation index to fresh islets, whereas only hCell- and MM1-cultured islets demonstrated recovery of C-peptide content and insulin release.
Media choice before transplant can influence islet quality, even when culture periods are short. Miami MM1 and hCell media may provide better islet protection than alternative media.
人类胰岛移植方案通常在移植前包含一段短暂的胰岛培养期。移植前胰岛培养的最佳培养基尚不清楚。
我们比较了四种为人类胰岛配制的无血清培养基：迈阿密（MM1）、孟菲斯（M-SFM）、埃德蒙顿（EDM）和hCell OCZEM-SF/AF（hCell）。从单个纯度>80%的人类胰腺中分离出的胰岛，使用所列培养基以2500个胰岛当量（IE）的份数进行培养。7天后，将每个2500-IE的份数移植到链脲佐菌素诱导的糖尿病rag1小鼠的肾包膜下（每组n = 4）。通过监测血清葡萄糖、刺激C肽释放和葡萄糖耐量试验对小鼠进行评估。分离后立即移植的胰岛份数（n = 4只小鼠）作为对照。在第0天和第3天评估体外胰岛功能，包括胰岛素释放（静态葡萄糖刺激后）、总细胞C肽含量、细胞计数和活力。
移植后所有小鼠组的血糖控制均得到改善，但只有在MM1中培养的胰岛移植与新鲜胰岛在统计学上无差异。与新鲜胰岛相比，MM1和hCell培养的胰岛移植显示出更好的葡萄糖耐量；四个组之间的C肽释放相似。在体外，只有在MM1中培养的胰岛与新鲜胰岛具有相似的刺激指数，而只有hCell和MM1培养的胰岛表现出C肽含量和胰岛素释放的恢复。
即使培养期较短，移植前的培养基选择也会影响胰岛质量。迈阿密MM1和hCell培养基可能比其他培养基提供更好的胰岛保护。
