Porcine Neonatal Pancreatic Cell Clusters in Tissue Culture: Benefits of Serum and Immobilization in Alginate Hydrogel1

Tatarkiewicz, Krystyna; García, Maritza; López-Ávalos, María Dolores; Bonner-Weir, Susan; Weir, Gordon C.

doi:10.1097/00007890-200106150-00007

Cited by 22 publications

(17 citation statements)

References 39 publications

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“…As expected due to the limitations on cell mobility imposed by the alginate network, no such aggregation was seen for the encapsulated cell clusters. This is in line with previous work where alginate beads were shown to be efficient in preventing aggregation of porcine neonatal pancreatic cell clusters [45]. The encapsulated clusters were found to be of similar size at both day 14 and 46 after encapsulation, ranging from 50 to 400 µm.…”

Section: Screening Of Encapsulation Materials For Efficiency As Facilsupporting

confidence: 91%

Culture of hESC‐derived pancreatic progenitors in alginate‐based scaffolds

Formo

Cho

Vallier

et al. 2015

J Biomedical Materials Res

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The effect of alginate-based scaffolds with added basement membrane proteins on the in vitro development of hESC-derived pancreatic progenitors was investigated. Cell clusters were encapsulated in scaffolds containing the basement membrane proteins collagen IV, laminin, fibronectin, or extracellular matrix-derived peptides, and maintained in culture for up to 46 days. The cells remained viable throughout the experiment with no signs of central necrosis.Whereas non-encapsulated cells aggregated into larger clusters, some of which showed signs of morphological changes and tissue organization, the alginate matrix stabilized the cluster size and displayed more homogeneous cell morphologies, allowing culture for long periods of time. For all conditions tested, a stable or declining expression of insulin and PDX1 and an increase in glucagon and somatostatin over time indicated a progressive reduction in beta cellrelated gene expression. Alginate scaffolds can provide a chemically defined, xeno-free and easily scalable alternative for culture of pancreatic progenitors. Although no increase in insulin and PDX1 gene expression after alginate-immobilized cell culture was seen in this study, further optimization of the matrix physicochemical and biological properties and of the medium composition may still be a relevant strategy to promote the stabilization or maturation of stem cell-derived beta cells.

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Section: Screening Of Encapsulation Materials For Efficiency As Facilsupporting

confidence: 91%

Culture of hESC‐derived pancreatic progenitors in alginate‐based scaffolds

Formo

Cho

Vallier

et al. 2015

J Biomedical Materials Res

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“…10B). This level of long-term cell survival was comparable or superior to our previous experience with neonatal pancreatic porcine cells (Hoesli et al, 2009) and other reports of 20-30% primary cell survival after 10 days of alginate-immobilized culture (Tatarkiewicz et al, 2001;Yang et al, 2002). Insulin expression increased significantly for all conditions over the 10 days of serum-free culture by 4 AE 2 fold for adherent cultures, 21 AE 5 fold for suspension cultures and 67 AE 32 fold (ranging from 11-to 159-fold) for the immobilized cultures in emulsion-generated alginate beads (Fig.…”

Section: Culture Of Primary Islet-depleted Pancreatic Cells In Emulsisupporting

confidence: 86%

“…Since the islets used to treat one patient are usually procured from two to three cadaveric donors, widespread access to this treatment would require $100-fold increase in tissue availability (Ricordi, 2003). To satisfy this demand, glucose-responsive insulin-producing cells could be procured from differentiated ES cells (D'Amour et al, 2006), differentiated adult pancreatic progenitor cells (Xu et al, 2008), expanded b cells (Efrat, 2008;Tsang et al, 2007), or xenogeneic tissue (Dufrane and Gianello, 2008;Living Cell Technologies, 2010;Tatarkiewicz et al, 2001). All of these cell options could benefit from alginate immobilization as a method to increase the insulin content of the tissue in vitro and/or as part of the final product to protect the graft after transplantation.…”

Section: Introductionmentioning

confidence: 99%

Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation

Hoesli¹,

Raghuram²,

Kiang³

et al. 2010

Biotech & Bioengineering

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Alginate has been used to protect transplanted pancreatic islets from immune rejection and as a matrix to increase the insulin content of islet progenitor cells. The throughput of alginate bead generation by the standard extrusion and external gelation method is limited by the rate of droplet formation from nozzles. Alginate bead generation by emulsion and internal gelation is a scaleable alternative that has been used with biological molecules and microbial cells, but not mammalian cells. We describe the novel adaptation of this process to mammalian cell immobilization. After optimization, the emulsion process yielded 90 ± 2% mouse insulinoma 6 (MIN6) cell survival, similar to the extrusion process. The MIN6 cells expanded at the same rate in both bead types to form pseudo-islets with increased glucose stimulation index compared to cells in suspension. The emulsion process was suitable for primary pancreatic exocrine cell immobilization, leading to 67 ± 32 fold increased insulin expression after 10 days of immobilized culture. Due to the scaleability and broad availability of stirred mixers, the emulsion process represents an attractive option for laboratories that are not equipped with extrusion-based cell encapsulators, as well as for the production of immobilized or encapsulated cellular therapeutics on a clinical scale.

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“…The small subunits (called pancreatic cell clusters) were able to reverse hyperglycemia after transplantation. The authors were interested in methods to block aggregation of the cells for transplantation (21), and they did not directly test how the pancreatic cell clusters compared with large islets. There have been other reports (4, 11) that indicated differences in the size of the islets.…”

Section: Discussionmentioning

confidence: 99%

Small rat islets are superior to large islets in in vitro function and in transplantation outcomes

Macgregor¹,

Williams²,

Tong³

et al. 2006

American Journal of Physiology-Endocrinology and Metabolism

155

162

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Barriers to the use of islet transplantation as a practical treatment for diabetes include the limited number of available donor pancreata. This project was designed to determine whether the size of the islet could influence the success rate of islet transplantations in rats. Islets from adult rats were divided into two groups containing small (diameter Ͻ125 m) or large (diameter Ͼ150 m) islets. An average pancreas yielded three times more small islets than large. Smaller islets were ϳ20% more viable, with large islets containing a scattered pattern of necrotic and apoptotic cells or central core cell death. Small islets in culture consumed twice as much oxygen as large islets when normalized for the same islet equivalents. In static incubation, small islets released three times more insulin under basal conditions than did large islets. During exposure to high glucose conditions, the small islets released four times more insulin than the same islet equivalencies of large islets, and five times more insulin was released by the small islets in response to glucose and depolarization with K ϩ . Most importantly, the small islets were far superior to large islets when transplanted into diabetic animals. When marginal islet equivalencies were used for renal subcapsular transplantation, large islets failed to produce euglycemia in any recipient rats, whereas small islets were successful 80% of the time. The results indicate that small islets are superior to large islets in in vitro testing and for transplantation into the kidney capsule of diabetic rats. islet transplant; insulin secretion; viability THE RISE IN CASES OF DIABETES MELLITUS in the United States has been called an epidemic. It is the third leading cause of death by disease and rivals heart disease and cancer as a major killer of United States citizens. For unexplained reasons the occurrence of type 1 diabetes is increasing worldwide, and the age of onset has decreased by 3-5 yr over the past decade so that many children now develop diabetes before entering school. The result is that more people with diabetes will spend a larger percentage of their life at risk for developing the chronic complications related to type 1 diabetes. Because the risk for development of most of the chronic complications associated with diabetes is related to glycemic control, significant attention is directed toward novel therapies, such as islet transplantation, to improve glycemic control.Islet transplants in humans were first attempted in the 1980s (7, 8). Initial success rates for human islet transplantation were disappointing, with only 5% of patients receiving transplants achieving partial function (19). Amid the failures were isolated success stories of individuals achieving prolonged reversal of their diabetes for a 1-to 2-yr period (19), which encouraged researchers to continue this approach for the treatment of diabetes. In 2000, islet transplantations were performed successfully on seven patients with diabetes by using a suppression regimen that omitted glucocorticoid...

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Porcine Neonatal Pancreatic Cell Clusters in Tissue Culture: Benefits of Serum and Immobilization in Alginate Hydrogel1

Cited by 22 publications

References 39 publications

Culture of hESC‐derived pancreatic progenitors in alginate‐based scaffolds

Culture of hESC‐derived pancreatic progenitors in alginate‐based scaffolds

Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation

Small rat islets are superior to large islets in in vitro function and in transplantation outcomes

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