Detection of porcine umbilical cord matrix stem cells in the intestine and other organs after oral and intraperitoneal administration to allogeneic recipients

Abstract

Umbilical cords matrix stem cells (UCs) have been characterized most thoroughly in humans (HUCs) and are considered to have great promise for regenerative medicine and cell-based therapy. Although UCs were first identified in pigs the description of porcine UCs (PUCs) is limited. Here we reported some standard mesenchymal stem cell characteristics for PUCs. Development of knowledge about PUCs is useful because the pig is a valuable biomedical model for humans and the species is an important human food source. PUCs were isolated from Wharton’s jelly using an explant technique. They attached on the plastic and showed fibroblast-like morphology. Immunophenotype analysis showed they are positive for CD44, CD90 and CD105 and negative for CD31, CD45 and SLA-DR. Under specific in vitro conditions, PUCs were differentiated to adipocytes, chondrocytes and osteocytes. The growth curve of PUCs exhibited a lag phase, log phase and doubling time of 24, 60 and 13.8 hour respectively. Engraftment potential of allogeneic PUCs administered orally and intraperitoneally (IP) was evaluated. Newborn, 1-day, 1-week, 2-week and 3-week old pigs were administered a dose of fluorescently labeled PUCs (1.1x107 cells/kg body weight) and their tissue incorporation were evaluated using confocal microscopy with confirmation by PCR to detect SRY gene, the Y-chromosome gene of male PUCs in female recipients. One week after PUCs administration, they were found mostly in the gastrointestinal tract and abdominal organs after either oral or intraperitoneal transplantation. The intestinal mucosa layer around the base of villi and intestinal crypts was the main location. PUCs were also detected in thoracic organs, muscle and bone marrow. Additionally, PKH26-labeled fibroblasts labeled were detected in recipient intestine 1 week after IP injection. Donor cells were not found in blood at one week post transplantation. When recipients were sacrificed at 6 h after IP injection PKH26-labeled PUCs were found mostly in omentum and diaphragm by PCR. It is likely these are the primary sites for donor cells in the peritoneal cavity to enter the circulation. Fluorescent in situ hybridization (FISH), using an SRY probe and PCR, demonstrated the PUCs isolated from recipient intestines by enzymatic digestion. Therefore, transplanted PUCs were recovered from the intestinal mucosa and were viable and able to proliferate in vitro.