Functional restoration of ex vivo model of pylorus: Co-injection of neural progenitor cells and interstitial cells of Cajal

Abstract

Transplantation of neural stem cells is a promising approach in treatment of intestinal dysfunctionality. The interstitial cells of Cajal (ICCs) are also critical in conditions such as pyloric dysfunctionality and gastroparesis. The objective of this study was to replenish neurons and ICCs in a dysfunctional pylorus as cell-based therapy to restore functionality. ICCs and enteric neural progenitor cells (NPCs) were isolated from rat duodenum and transduced with fluorescent proteins. Rat pylorus was harvested, and an ex-vivo neuromuscular dysfunctional model was developed by selective ablation of neurons and ICCs via chemical treatments. Cellular repopulation and restoration of motility were assessed by immunohistochemistry, qPCR, and functional analysis after delivery of fluorescently tagged cells. Chemical treatment of pylorus resulted in significant depletion of ICCs (67%, P = .0024; n = 3) and neural cells (83%, P = .0012; n = 3). Delivered ICCs and NPCs survived and integrated with host muscle layers. Coinjection of ICCs with NPCs exhibited 34.4% (P = .0004; n = 3) and 61.0% (P = .0003; n = 3) upregulation of ANO1 and βIII tubulin, respectively. This regeneration resulted in the restoration of agonist-induced excitatory contraction (82%) and neuron evoked relaxation (83%). The functional studies with specific neuronal nitric oxide (NO) synthase blocker confirmed that restoration of relaxation was NO mediated and neuronally derived. The simultaneous delivery of ICCs observed 35.7% higher neuronal differentiation and functional restoration compared with injection of NPCs alone.Injected NPCs and ICCs integrated into the dysfunctional ex vivo pylorus tissues and restored neuromuscular functionality. The co-transplantation of NPCs and ICCs can be used to treat neurodegenerative disorders of the pylorus.

Structural Relationships Between Interstitial Cells of Cajal and Smooth Muscle Cells/Nerve Fibers in the Gastric Muscularis Mucosae of Chinese Giant Salamander

“…The illustration of ICC locations and their structural relationships with surrounding cell types will help us to better understand the physiological roles of ICC in motility and digestion of stomach, and get an insight into the etiology and pathogenesis of amphibian stomach motility disorders. In future, ICC will be a promising target cell and used potentially in clinical for the treatment of the abnormal motility of the GI tract by various methods, including ICC transplantation (Dadhich & Bitar, 2020).…”

An Engineered Living Intestinal Muscle Patch Produces Macroscopic Contractions that can Mix and Break Down Artificial Intestinal Contents

“…Various engineering strategies have regenerated smooth muscle that shows nonspontaneous contraction. [31][32][33][34][35] Several other pieces of research show that primary intestinal organoids, [36,37] primary cells isolated from the intestinal muscle layers (IMCs), [38,39] or primary intestinal muscle strips [40] can exert spontaneous (in some cases, rhythmic [38,40] ) contractions but only for no more than 14 days during in vitro culture. Contractile activity of IMCs can be extended to over ten weeks in vitro when they are cocultured with feeder cells.…”

A preview of selected articles

“…In the first of our Featured Articles this month from STEM CELLS Translational Medicine, Dadhich and Bitar employ an ex vivo rat model of gastroparesis to demonstrate how the administration of neural progenitor cells (NPCs) can prompt optimal restoration of functionality when accompanied by specialized gastrointestinal tract interstitial cells. 4 In a Related Article published in STEM CELLS, Lin et al discovered that the administration of in vitro preconditioned MSCs represent a promising therapy for gastrointestinal dysfunctions brought on by enteric nervous system injury or neurodegenerative disorders. 5 The efficient homing of hematopoietic stem and progenitor cells (HSPCs) to the bone marrow can significantly influence hematopoietic repopulation in patients undergoing hematopoietic stem cell transplantation (HSCT), as studies have established that the majority of HSPCs become trapped or "tethered" in various nonhematopoietic organs.…”

“…although the ability of said cells to migrate, proliferate, and generate functional neurons remained unknown. Now, a new STEM CELLS Translational Medicine article from Prabhash Dadhich and Khalil N. Bitar (Wake Forest Institute forRegenerative Medicine, Winston-Salem, North Carolina) explores the cotransplantation of adult NPCs and interstitial cells of Cajal (ICCs)12 in an ex vivo gastroparesis model as an optimized therapeutic approach 4. The authors first ablated neurons and ICCs in the rat pylorus to generate gastroparesis-associated neuromuscular problems and then codelivered ICCs and NPCs into the injured area.…”