Generation of mesenchymal stromal cells from urine-derived iPSCs of pediatric brain tumor patients

Abstract

Human induced pluripotent stem cells (iPSCs) provide a virtually inexhaustible source of starting material for next generation cell therapies, offering new opportunities for regenerative medicine. Among different cell sources for the generation of iPSCs, urine cells are clinically relevant since these cells can be repeatedly obtained by non-invasive methods from patients of any age and health condition. These attributes encourage patients to participate in preclinical and clinical research. In particular, the use of urine-derived iPSC products is a convenient strategy for children with brain tumors, which are medically fragile patients. Here, we investigate the feasibility of using urine samples as a source of somatic cells to generate iPSC lines from pediatric patients with brain tumors (BT-iPSC). Urinary epithelial cells were isolated and reprogrammed using non-integrative Sendai virus vectors harboring the Yamanaka factors KLF4, OCT3/4, SOX2 and C-MYC. After reprogramming, BT-iPSC lines were subject to quality assessment and were compared to iPSCs obtained from urine samples of non-tumor pediatric patients (nonT-iPSC). We demonstrated that iPSCs can be successfully derived from a small volume of urine obtained from pediatric patients. Importantly, we showed that BT-iPSCs are equivalent to nonT-iPSCs in terms of morphology, pluripotency, and differentiation capacity into the three germ layers. In addition, both BT-iPSCs and nonT-iPSCs efficiently differentiated into functional mesenchymal stem/stromal cells (iMSC) with immunomodulatory properties. Therefore, this study provides an attractive approach to non-invasively generate personalized iMSC products intended for the treatment of children with brain tumors.

Urine-derived stem cells: Promising advancements and applications in regenerative medicine and beyond

“…In neurologically relevant studies, USCs have proven valuable in neuroprotection and therapy. Exosome-derived miR-21-5p from USCs has been shown to attenuate the symptoms of Rett syndrome by modulating the EPha4/TEK axis [ 44 , 90 ]. USC-exos carrying ANGPTL3 can enhance functional recovery of the spinal cord after injury by promoting angiogenesis [ 91 , 92 ].…”

“…These BT-iPSCs exhibited comparable morphology, pluripotency, and the ability to differentiate into the three blastomeres when compared to iPSCs obtained from paediatric patients without tumors (non-T-iPSCs). This result and technique offer a nearly limitless source of starting material for cellular therapeutics and introduce a new perspective for regenerative medicine [ 44 ]. Furthermore, genetically engineered induced cells offer models for understanding the basis of rare diseases, including those associated with abnormal nerve cell development such as Hemophilia A (HA), phenylketonuria (PKU), and Down syndrome (DS).…”