Determination of the potential of induced pluripotent stem cells to differentiate into mouse nucleus pulposus cells in vitro

Abstract

ABSTRACT. We determined the potential for induced pluripotent stem (iPS) cells to differentiate into nucleus pulposus (NP)-like cells in mice. iPS cells were generated from tail-tip fibroblasts. We used a pellet culture model with the aim of determining the applicability of iPS cell-based therapy to intervertebral disc degeneration (IVD). The cell pellet was cultured in an NP cell basal medium comprising Dulbecco's modified Eagle's medium supplemented with transforming growth factor beta 1, dexamethasone, ascorbate-2-phosphate, and 1% ITS-Premix. The pellet was evaluated by quantitative reverse transcription polymerase chain (2015) reaction, immunohistochemical staining, and biochemical composition. The differentiation of iPS cells into NP cells was demonstrated by the protein and mRNA expression levels of proteoglycan, collagen II, aggrecan, and CD24. Furthermore, increased hydroxyproline content and dimethylmethylene blue staining demonstrated that the collagen II and glycosaminoglycan content in the NP cells increased with time. We have shown that cultured mouse iPS cells can be induced to differentiate into NP cells. Such proofof-concept opens up the possibility of producing patient-specific NP cells in a relatively simple and straightforward manner with high efficiency. We are confident that such cells could be immediately useful for the study of IVD disease.

Efficacy of administered mesenchymal stem cells in the initiation and co‐ordination of repair processes by resident disc cells in an ovine (Ovis aries) large destabilizing lesion model of experimental disc degeneration

“…Caplan proposed that stem cells should be renamed Medicinal Signaling Cells to more accurately reflect how they home in on injured or diseased tissue sites secreting bioactive factors with immunomodulatory and trophic properties which direct the resident cells to undertake the tissue repair process, this may happen long after the MSCs were present in the defect site . The use of MSCs to undertake IVD repair/regeneration is an area which shows much promise with many publications appearing in the literature dealing with animal model based laboratory studies, preclinical studies, clinical trials (Tables and ) and reviews advocating the use of MSCs for disc repair/regeneration …”

“…Medicinal Signaling Cells to more accurately reflect how they home in on injured or diseased tissue sites secreting bioactive factors with immunomodulatory and trophic properties which direct the resident cells to undertake the tissue repair process, this may happen long after the MSCs were present in the defect site. 27 The use of MSCs to undertake IVD repair/regeneration is an area which shows much promise [28][29][30] with many publications appearing in the literature dealing with animal model based laboratory studies, [31][32][33][34][35][36] preclinical studies, 37,38 clinical trials 39 (Tables 1 and 2) and reviews advocating the use of MSCs for disc repair/regeneration. 28,29,[45][46][47][48][49][50][51] Most pre-clinical studies have treated IVDs with MSCs acutely after induction of disc degeneration, and no direct studies on the relative efficacy of MSCs in different stages of the disease process have been evaluated.…”

The generation and functional characterization of induced pluripotent stem cells from human intervertebral disc nucleus pulposus cells

“…Chen et al differentiated iPSCs into the NP-like cell phenotype with native NP tissue culture media under hypoxic conditions, the enriched CD24+ fraction of iPSCs was directed toward an NP-like phenotype without growth factors [21]. Feng et al differentiated mouse pluripotent stem cells into functional NP cells with transforming growth factor beta 1 (TGF-β1) [22]. Both protocols resulted in the expression of NP cell lineage markers and NP functional characteristics.…”

Novel nano-microspheres containing chitosan, hyaluronic acid, and chondroitin sulfate deliver growth and differentiation factor-5 plasmid for osteoarthritis gene therapy

“…Chondrocytes were cultured for one week after transfection and the cells were collected for biochemical assays. The assay method was as previously reported (Liu et al, 2015). Briefly, cells were digested and then washed with PBS three times for further assay.…”