Pancreatic Endocrine and Exocrine Cell Ontogeny From Renal Capsule–transplanted Embryonic Stem Cells in Streptozocin-injured Mice

Abstract

S U M M A R Y In this study, we describe pancreatic cell ontogeny in renal capsule-transplanted embryonic stem cells (ES) after injury by streptozocin (STZ), showing pancreatogenesis in situ. Seven-week-old female BALB/c nude mice were treated with either a single 175-or 200-mg/ kg STZ dose, a regimen that induces substantial b-cell damage without overt hyperglycemia, and transplanted 24 hr later with 1 3 10 5 ES. Immunohistochemistry was performed on ES tissue at 15, 21, and 28 days after transplantation using antibodies against stage-and lineagespecific pancreatic markers. After 21 days, PDX-11 pancreatic foci first appeared in the renal capsule and expressed both amylase and endocrine hormones (insulin, glucagon, and somatostatin). These foci increased in size by day 28 because of acinar and duct cell proliferation, whereas endocrine cells remained non-dividing, and made up 2-4% of ES tumor volume. PDX-1, Nkx6.1, Ngn3, and ISL-1 protein localization patterns in pancreatic foci were comparable with embryonic pancreatogenesis. A prevalence of multihormonal endocrine cells, a characteristic of adult b-cell regeneration, indicated a possible divergence from embryonic islet cell development. The results indicate that b-cell damage, without overt hyperglycemia, induces a process of fetal-like pancreatogenesis in renal capsule-transplanted ES, leading to b-cell neogenesis. (J Histochem Cytochem 56:33-44, 2008)

Embryonic Stem Cell Transplantation Correlates With Endogenous Neurogenin 3 Expression and Pancreas Regeneration in Streptozotocin-injured Mice

“…Typically, the pancreas-localized ES cells existed as small clusters adherent to the interstitium and adjacent to sites of neogenesis. Contrary to our expectations based on former data (Kodama et al 2008), the ES cells themselves were never identified as components of normal ductal, exocrine, or endocrine tissue within the pancreas. However, areas of regeneration (PDX-1 1 / Ngn3 1 ductal tissue) and ES cell-derived cells were closely associated, a typical example of which is shown in Figure 4.…”

“…Non-STZ-treated mice received 300 ml citrate buffer alone. Twenty-four hr after STZ treatment, ES cells were transplanted beneath the left renal capsule as described previously (Kodama et al 2008). PBS was dispensed under the renal capsule for sham transplantation.…”

“…STZ was administered at 200 mg/kg, which in our hands typically induces overt hyperglycemia: .200 mg/dl blood glucose concentration for three consecutive readings in ,50% of animals (Kodama et al 2008). Hence, most of the animals were euglycemic at the time of sacrifice (see supplementary Table 1).…”

Stem Cell Therapy for Islet Regeneration

“…Twenty-one days posttransplantation, PDX-1+ pancreatic foci appeared in the renal capsule, which expressed exocrine enzymes (amylase) and endocrine hormones (insulin, glucagon, and somatostatin). These multi-hormonal endocrine cells, a characteristic of beta cell regeneration, suggested possible divergence from embryonic islet cell development (Kodama et al, 2008). In another study, Kodama et al (2009) showed that transplanted ESCs could migrate into the injured pancreas.…”

“…Many different types of stem cells have been tested using different methods. Cells can be grafted underneath the kidney capsule (Rackham et al, 2011;Figliuzzi et al, 2009;Ito et al, 2010;Kodama et al, 2009;Kodama et al, 2008;Zhang et al, 2010;Ohmura et al, 2010;Xiao et al, 2008;Berman et al, 2010), delivered via intra-peritoneal injection (Boroujeni et al, 2011;Chandra et al, 2009;Koya et al, 2008;Shao et al, 2011;Phuc et al, 2011; or intra-portally (Shyu et al, 2011;Trivedi et al, 2008;Wu et al, 2007;Longoni et al, 2010;Itakura et al, 2007), grafted into the liver (Chao et al, 2008;Zhu et al, 2009;Xu et al, 2007;Wang et al, 2010) or injected into the tail vein (Dinarvand et al, 2010;Koblas et al, 2009;Kajiyama et al, 2010;Jurewicz et al, 2010) (Fig. 3).…”