Int J Stem Cells.  2018 Nov;11(2):205-215. 10.15283/ijsc18002.

Effect of Stem Cells and Gene Transfected Stem Cells Therapy on the Pancreas of Experimentally Induced Type 1 Diabetes

• Zickri MB
• Aboul-Fotouh GI
• Omar AI
• El-Shafei
• Reda AM

• KMID: 2429553
• DOI: http://doi.org/10.15283/ijsc18002

Abstract

BACKGROUND AND OBJECTIVES
Insulin secretion entirely depends on Ca²âº influx and sequestration into endoplasmic reticulum (ER) of β-cells, performed by Sarco-ER Ca²âº-ATPase 2b (SERCA2b). In diabetes, SERCA2b is decreased in the β-cells leading to impaired intracellular Ca²âº homeostasis and insulin secretion. Adipose mesenchymal stem cells (AMSCs) play a potential role in transplantation in animal models. The present study aimed at investigating and comparing the therapeutic effect of non-transfected AMSCs and SERCA2b gene transfected AMSCs on the pancreas of induced diabetes type 1 in rat.
METHODS AND RESULTS
58 adult male albino rats were divided into: Donor group: 22 rats, 2 for isolation, propagation and characterization of AMSCs and SERCA2b transfected AMSCs, in addition 20 for isolated islet calcium level assessment. Group I (Control Group): 6 rats, Group II (Diabetic Group): 10 rats, 50 mg streptozotocin (STZ) were injected intraperitoneal (IP), Group III (AMSCs Group): 10 rats, 1×10⁶ AMSCs were injected intravenous and Group IV (SERCA2b transfected AMSCs Group): 10 rats, 1×10⁶SERCA2b transfected AMSCs were injected as in group III. Groups I, II, III and IV were sacrified 3 weeks following confirmation of diabetes. Serological, histological, morphometric studies and quantitative polymerase chain reaction (qPCR) were performed. Nuclear, cytoplasmic degenerative and extensive fibrotic changes were detected in the islets of group II that regressed in groups III and IV. Isolated islet calcium, blood glucose, plasma insulin and qPCR were confirmative.
CONCLUSIONS
AMSCs and SERCA2b gene transfected AMSCs therapy proved definite therapeutic effect, more obvious in response to SERCA2b gene transfected AMSCs.

Keyword

AMSCs; SERCA2b; Type 1 diabetes; Pancreas

MeSH Terms

Adult
Animals
Blood Glucose
Calcium
Cytoplasm
Endoplasmic Reticulum
Homeostasis
Humans
Insulin
Male
Mesenchymal Stromal Cells
Models, Animal
Pancreas*
Plasma
Polymerase Chain Reaction
Rats
Stem Cells*
Streptozocin
Tissue Donors
Blood Glucose
Calcium
Insulin
Streptozocin

Figure

• 

  Fig. 1 Showing: (A) CD44 +ve spindle cells (arrows). (B) CD34 −ve immunoreactivity (arrows) (Phase contrast microscopy×100). (C) Immunophenotyping of AMSCs 96% are +ve for CD44 (Flow Cytometry). (D) Transfected cells appearing mostly as spindle fluroscent labeled cells (GFP×100). (E) Immunophenotyping of transfected AMSCs showed 93.3% of the cells GFP labeled (Flow Cytometry).

• 

  Fig. 2 Showing: (A) the islet cells exhibiting pale nuclei (N) and secretory granules (g) in group I. (B) a shrunken islet (s) exhibiting fibroblasts (f) and few cells with absent secretory granules in group II. (C) condensed nuclear chromatin (cc) of an islet cell, multiple pale nuclei (N) and secretory granules (g) in group III. (D) condensed nuclear chromatin (cc) of an islet cell, pale nuclei (N) and secretory granules (g) in most islet cells of group IV (Toluidine blue, ×1000).

• 

  Fig. 3 Showing: (A) fine collagen fibers between the acini (arrows) and among the islet cells (arrowheads) in group I. (B) multiple collagen fibers between the acini (arrows) and among the islet cells (arrowheads) in group II. (C) minimal increase in the collagen content (arrow) around a vessel (V) and fine collagen fibers among the islet cells (arrowheads) in group III. (D) fine collagen fibers between the acini (arrow) and among the islet cells (arrowhead) in group IV (Masson’s Trichrome, ×200).

• 

  Fig. 4 Showing: (A) some fluorescent labeled cells (arrows) among pancreatic acini and an islet (rectangle) in group III (PKH26, ×100). (B) Few fluorescent labeled cells (arrows) among pancreatic acini and an islet (rectangle) in group IV (GFP, ×100).

• 

  Fig. 5 Showing: (A) −ve IE in group I. (B) multiple +ve spindle cells among islet cells (arrows) and acini (thick arrow) in group II. (C) multiple +ve cells among islet cells (arrows) and acini (thick arrow) in group III. (D) some +ve cells among islet cells (arrow) and acini (thick arrow) in group IV (CD44 immunostaining, ×400).

• 

  Fig. 6 Showing: +ve IE among (arrowhead): (A) most of islet cells in group I. (B) few cells in a shrunken islet of group II. (C) multiple islet cells in group III. (D) most islet cells in group IV (Insulin immunostaining, ×400).

Reference

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