Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, Hanoi, Vietnam.
In Vitro Cell Dev Biol Anim. 2011 Jan;47(1):54-63. doi: 10.1007/s11626-010-9356-5. Epub 2010 Nov 17.
Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are multipotent cells. They are able to differentiate into functional cells from not only mesoderm but also endoderm. Many researches showed that cells derived from fresh human UCB could transdifferentiate into insulin-secreting cells. In this study, transdifferentiating potential of cryopreserved human UCB-derived MSCs into insulin-secreting cell was investigated. Fresh human UCB was enriched the mononuclear cells by Ficoll-Paque centrifugation. The mononuclear cell population was cryopreserved in cryo-medium containing Iscove's modified Dulbecco's media (IMDM) with 10% DMSO at -196°C for 1 yr. After thawing, mononuclear cells were cultured to isolate MSCs in medium IMDM with 20% FBS supplemented with growth factors. At the fifth passages, MSCs were confirmed by flow cytometry about expression of CD13, CD14, CD34, CD45, CD166, and HLA-DR markers; after that, they were induced to differentiate into adipocytes and osteoblasts. After inducing with specific medium for islet differentiation, there were many clusters of cell like islet at day 14-28. Using real-time reverse transcription polymerase chain reaction (RT-PCR) to analyze the expression of functional genes, the result showed that Nestin, Pdx-1, Ngn3, Ils-1, Pax6, Pax4, Nkx2.2, Nkx6.1, Glut-2, Insulin genes expressed. The results showed that MSCs derived from banked cord blood can differentiate into functional pancreatic islet-like cells in vitro. If human MSCs, especially MSCs from banked cord blood of diabetes patients themselves can be isolated, proliferated, differentiated into functional pancreatic islet-like cells, and transplanted back into them (autologous transplantation), their high-proliferation potency and rejection avoidance will provide one promising therapy for diabetes.
脐带血（UCB）衍生的间充质干细胞（MSCs）是多能细胞。它们不仅可以从中胚层分化为功能性细胞，还可以从内胚层分化为功能性细胞。许多研究表明，来自新鲜人 UCB 的细胞可以转分化为分泌胰岛素的细胞。在这项研究中，研究了冷冻保存的人 UCB 衍生 MSC 向胰岛素分泌细胞的转分化潜能。通过 Ficoll-Paque 离心从新鲜人 UCB 中富集单核细胞。将单核细胞群体在含有 IMDM 中 10% DMSO 的冷冻培养基中于-196°C 冷冻保存 1 年。解冻后，将单核细胞在含有 20% FBS 的 IMDM 培养基中培养以分离 MSC，并补充生长因子。在第五代时，通过流式细胞术确认 MSC 表达 CD13、CD14、CD34、CD45、CD166 和 HLA-DR 标志物；之后，将其诱导分化为脂肪细胞和成骨细胞。在用特定的胰岛分化培养基诱导后，第 14-28 天会出现许多胰岛样细胞簇。使用实时逆转录聚合酶链反应（RT-PCR）分析功能基因的表达，结果表明巢蛋白、PDX-1、Ngn3、胰岛素样生长因子-1、Pax6、Pax4、Nkx2.2、Nkx6.1、Glut-2、胰岛素基因表达。结果表明，来自银行脐带血的 MSC 可以在体外分化为功能性胰岛样细胞。如果可以分离、增殖、分化来自糖尿病患者自身银行脐带血的人 MSC 为功能性胰岛样细胞，并将其移植回体内（自体移植），它们的高增殖能力和避免排斥反应将为糖尿病提供一种有前途的治疗方法。
