Dental Pulp Cells for Induced Pluripotent Stem Cell Banking

Abstract

Defined sets of transcriptional factors can reprogram human somatic cells to induced pluripotent stem (iPS) cells. However, many types of human cells are not easily accessible to minimally invasive procedures. Here we evaluated dental pulp cells (DPCs) as an optimal source of iPS cells, since they are easily obtained from extracted teeth and can be expanded under simple culture conditions. From all 6 DPC lines tested with the conventional 3 or 4 reprogramming factors, iPS cells were effectively established from 5 DPC lines. Furthermore, determination of the HLA types of 107 DPC lines revealed 2 lines homozygous for all 3 HLA loci and showed that if an iPS bank is established from these initial pools, the bank will cover approximately 20% of the Japanese population with a perfect match. Analysis of these data demonstrates the promising potential of DPC collections as a source of iPS cell banks for use in regenerative medicine.

<b>Induction of neural crest cells from human dental pulp-derived induced pluripotent stem </b><b>cells </b>

“…Therefore, NCCs could represent a key cell source for tooth-tissue engineering. Induced pluripotent stem (iPS) cells are generated from a variety of somatic cells such as dermal fibroblasts (53,54), blood cells, (12) and dental pulp cells (42,55,59,62) by induction with several factors. iPS cells are able to differentiate into all three germ layers with large-scale expansion (12, 42, 53-55, 59, 62), bypassing the potential ethical problems of using embryonic stem (ES) cells.…”

“…Therefore, iPS cells offer the potential to derive patient-and lineage-specific cells for clinical applications and have emerged as a potential cell source for regenerative medicine and tissue engineering. Previous studies suggested that cell proliferation ability (42,59) or endogenous expression of reprogramming factors (10,55) in parent cells is positively related to the reprogramming efficiency. Human dental pulp cells as parent cells for iPS cells exhibit higher reprogramming efficiency than do human dermal fibroblasts (42,55) because they show rapid cell proliferation (42) and high expression of endogenous reprogramming factors, c-MYC and KLF4 (55), compared to human dermal fibroblasts.…”

“…Previous studies suggested that cell proliferation ability (42,59) or endogenous expression of reprogramming factors (10,55) in parent cells is positively related to the reprogramming efficiency. Human dental pulp cells as parent cells for iPS cells exhibit higher reprogramming efficiency than do human dermal fibroblasts (42,55) because they show rapid cell proliferation (42) and high expression of endogenous reprogramming factors, c-MYC and KLF4 (55), compared to human dermal fibroblasts. Since dental pulp cells are readily obtained from extracted teeth and can proliferate well in normal culture conditions (55), no further procedures are necessary in regard to the donors.…”

“…Human dental pulp cells as parent cells for iPS cells exhibit higher reprogramming efficiency than do human dermal fibroblasts (42,55) because they show rapid cell proliferation (42) and high expression of endogenous reprogramming factors, c-MYC and KLF4 (55), compared to human dermal fibroblasts. Since dental pulp cells are readily obtained from extracted teeth and can proliferate well in normal culture conditions (55), no further procedures are necessary in regard to the donors. iPS plemented with 0.5× N-2 (Thermo Fisher Scientific), 0.5× B-27 (Thermo Fisher Scientific), 5 μg/mL insulin (Sigma-Aldrich), 20 ng/mL bFGF (Wako), 20 ng/ mL epidermal growth factor (EGF; Sigma-Aldrich), 50 U/mL penicillin, and 50 μg/mL streptomycin, as in previous studies (4,39,43).…”

Tissue Engineering for Tissue and Organ Regeneration

“…The biological constraints, which become better characterized as research progresses, will necessarily restrict the number of protocols and strategies. Potential human cell sources have been reported (Gronthos et al, 2000;Miura et al, 2003), while banks for autologous cell sources are being developed (Arora et al, 2009;Tamaoki et al, 2010;Woods et al, 2009). The biological potentialities of the different cells types, which recently were identified from human sources, will have to be tested both in vitro and in vivo.…”