Differentiating of banked human umbilical cord blood-derived mesenchymal stem cells into insulin-secreting cells

Abstract

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.

Preliminary evaluation of intravenous infusion and intrapancreatic injection of human umbilical cord blood-derived mesenchymal stem cells for the treatment of diabetic mice

“…All procedures related to the handling of the blood samples were approved by the Ethical Committee of our university. MSCs from human umbilical cord blood were cultured according to previously published procedure (Phuc et al, 2011). Briefly, mononuclear cells (MNCs) were isolated from umbilical cord blood by centrifuging with Ficoll 1.077 (GE-Healthcare).…”

Stem Cell Therapy for Islet Regeneration

“…MSCs have been isolated from many different tissues, including bone marrow (Oyajobi et al. 1999;Majumdar et al, 2000;Prockop et al, 2001;Smith et al, 2004;Titorencu et al, 2007;Wolfe et al, 2008;Gronthos and Zannettino et al, 2008;Phadnis et al, 2011;Bao et al, 2011), adipose tissue (Katz et al, 2005;Baptista et al, 2009;Caviggioli et al, 2009;Baer et al, 2010;Bruyn et al, 2010;Estes et al, 2010;Tucker, Bunnell, 2011), peripheral blood (Kassis et al, 2006), umbilical cord blood (Erices et al, 2000;Rosada et al, 2003;Hutson et al, 2005;Reinisch et al, 2007;Bieback and Klüter et al, 2007;Perdikogianni et al, 2008;Zhang et al, 2011), banked umbilical cord blood (Phuc et al, 2011), umbilical cords (Cutler et al, 2010Farias et al, 2011), umbilical cord membranes (Deuse et al, 2010;Kita et al, 2010), umbilical cord veins (Santos et al, 2010), Wharton's jelly from the umbilical cord (Zeddou et al, 2010;Peng et al, 2011), placenta (Miao et al, 2006;Battula et al, 2007;Huang et al, 2009;Semenov et al, 2010;Pilz et al, 2011), decidua basalis (Macias et al, 2010;Lu et al, 2011), the ligamentum flavum , amniotic fluid (Feng et al, 2009;…”

“…In some studies, epidermal growth factor (EGF) was added to the culture medium during the IPC maturation phase in addition to nicotinamide. Currently, IPCs can be generated from MSCs obtained from human umbilical cord blood Parekh et al, 2009;Wang et al, 2010), banked human umbilical cord blood (Phuc et al, 2011), placenta , bone marrow Xie et al, 2009;Phadnis et al, 2011), menstrual blood , amniotic fluid (Trovato et al, 2009), Wharton's jelly (Chao et al, 2008;, amnion , and adipose tissue (Chandra et al, 2009). Other studies report the successful use of transgenesis to differentiate MSCs into IPCs, or upregulation of genes (mainly PDX-1 or betacellulin) related to signalling pathways that trigger this process (Karnieli et al, 2007;Li et al, 2007;Li et al, 2008;Hisanaga et al, 2008;Limbert and Seufert., 2009;Yuan et al, 2010;Paz et al, 2011).…”

Tissue Engineering for Tissue and Organ Regeneration

“…The identification of these critical factors is one focus of current research. Amnion epithelial cells Kadam et al, 2010a), and stem cells from liver ), wharton's jelly of umbilical cord (Chao et al, 2008), umbilical cord blood (Phuc et al, 2010), placenta (Kadam et al, 2010b), gall bladder (Sahu et al, 2009) etc were also differentiated to insulin producing cells. In every case the differentiated islets expressed islet specific proteins and were capable of secreting insulin in response to glucose stimulation, albeit less than native islets.…”