In Vivo Transplantation of Neurosphere-Like Bodies Derived from the Human Postnatal and Adult Enteric Nervous System: A Pilot Study

Abstract

Recent advances in the in vitro characterization of human adult enteric neural progenitor cells have opened new possibilities for cell-based therapies in gastrointestinal motility disorders. However, whether these cells are able to integrate within an in vivo gut environment is still unclear. In this study, we transplanted neural progenitor-containing neurosphere-like bodies (NLBs) in a mouse model of hypoganglionosis and analyzed cellular integration of NLB-derived cell types and functional improvement. NLBs were propagated from postnatal and adult human gut tissues. Cells were characterized by immunohistochemistry, quantitative PCR and subtelomere fluorescence in situ hybridization (FISH). For in vivo evaluation, the plexus of murine colon was damaged by the application of cationic surfactant benzalkonium chloride which was followed by the transplantation of NLBs in a fibrin matrix. After 4 weeks, grafted human cells were visualized by combined in situ hybridization (Alu) and immunohistochemistry (PGP9.5, GFAP, SMA). In addition, we determined nitric oxide synthase (NOS)-positive neurons and measured hypertrophic effects in the ENS and musculature. Contractility of treated guts was assessed in organ bath after electrical field stimulation. NLBs could be reproducibly generated without any signs of chromosomal alterations using subtelomere FISH. NLB-derived cells integrated within the host tissue and showed expected differentiated phenotypes i.e. enteric neurons, glia and smooth muscle-like cells following in vivo transplantation. Our data suggest biological effects of the transplanted NLB cells on tissue contractility, although robust statistical results could not be obtained due to the small sample size. Further, it is unclear, which of the NLB cell types including neural progenitors have direct restoring effects or, alternatively may act via ‘bystander’ mechanisms in vivo. Our findings provide further evidence that NLB transplantation can be considered as feasible tool to improve ENS function in a variety of gastrointestinal disorders.

Schizophrenia and Human Self-Domestication: An Evolutionary Linguistics Approach

“…SOX10 is found to be hypermethylated in the brain of SZ patients [Iwamoto et al, 2005] and carries singlenucleotide polymorphisms that have been related to the age of onset of clinical manifestations [Yuan et al, 2013]. RET and SOX10 are coexpressed as reliable markers in the enteric nervous system, although their functional interaction is not clear [Hetz et al, 2014]. In turn, FOXD3 is required during NC development and regulates dorsal mesoderm development in the zebrafish [Chang and Kessler, 2010].…”

Delivery of enteric neural progenitors with 5-HT4 agonist-loaded nanoparticles and thermosensitive hydrogel enhances cell proliferation and differentiation following transplantation in vivo

“…A number of recent studies have shown that ENS stem/progenitor cells (ENSCs) can be isolated, expanded in culture, and transplanted into intestinal tissue in vitro [4-6] and in vivo [7-9], including into animal models of enteric neuropathies [10-12]. However, the capacity of these cells to proliferate and migrate has been limited [12, 13], and may therefore be insufficient to restore gut function, emphasizing the need to engineer the cells and/or their microenvironment, and to optimize cell delivery methods, in order to maximize the success of cell replacement therapy.…”

“…Most studies to date have introduced neuronal precursor cells by laparotomy and direct injection into the gut wall [10, 11, 18-20]. Although some have demonstrated improved gut function following cell injection [10-12], there are significant drawbacks to this delivery method, such as substantial leakage from the puncture hole and poorly controllable cell spreading. These factors lead to inefficient cell engraftment, random cell distribution, and poor reproducibility.…”

Human and Murine Tissue-Engineered Colon Exhibit Diverse Neuronal Subtypes and Can Be Populated by Enteric Nervous System Progenitor Cells When Donor Colon Is Aganglionic

“…18 ENS progenitor cells have been transplanted in various in vitro and in vivo models and shown to differentiate into neurons and glia. [18][19][20][21][22][23][24][25][26][27][28][29] Early evidence indicates that these cells are functional and able to electrophysiologically integrate within the host's ENS. 28,29 In parallel, tissue-engineered intestine is a promising therapy for organ replacement from autologous cells.…”

“…[18][19][20][21][22][23][24][25][26][27][28][29] Early evidence indicates that these cells are functional and able to electrophysiologically integrate within the host's ENS. 28,29 In parallel, tissue-engineered intestine is a promising therapy for organ replacement from autologous cells. Our laboratory has developed tissue-engineered colon (TEC) from innervated human and rat colonic tissue digested into multicellular clusters termed organoid units (OU) and shown that it contains differentiated epithelium, smooth muscle, and glial cells, 30,31 which are all donor derived.…”