Derivation of iPSCs after Culture of Human Dental Pulp Cells under Defined Conditions

Abstract

Human dental pulp cells (hDPCs) are a promising resource for regenerative medicine and tissue engineering and can be used for derivation of induced pluripotent stem cells (iPSCs). However, current protocols use reagents of animal origin (mainly fetal bovine serum, FBS) that carry the potential risk of infectious diseases and unwanted immunogenicity. Here, we report a chemically defined protocol to isolate and maintain the growth and differentiation potential of hDPCs. hDPCs cultured under these conditions showed significantly less primary colony formation than those with FBS. Cell culture under stringently defined conditions revealed a donor-dependent growth capacity; however, once established, the differentiation capabilities of the hDPCs were comparable to those observed with FBS. DNA array analyses indicated that the culture conditions robustly altered hDPC gene expression patterns but, more importantly, had little effect on neither pluripotent gene expression nor the efficiency of iPSC induction. The chemically defined culture conditions described herein are not perfect serum replacements, but can be used for the safe establishment of iPSCs and will find utility in applications for cell-based regenerative medicine.

Priming with FGF2 stimulates human dental pulp cells to promote axonal regeneration and locomotor function recovery after spinal cord injury

“…Dental pulp was collected from the third molars of patients at Gifu University Hospital, with informed consent from each patient; dental pulp cells (DPCs) were prepared as previously reported 34 , 35 . Briefly, using a protocol approved by the Institutional Review Board of Gifu University, we collected normal human third molars at the Gifu University Medical Hospital after having obtained informed consent from each patient.…”

Mesenchymal Stem Cells Derived from Dental Pulp: A Review

“…Although these quality-control and biosecurity concerns are more theoretical than real because no such problems have been reported for transplants of DPSCs obtained from cultures that were expanded in serum-containing medium, in response to this risk, various methods of serum-free culture, using chemically defined materials as supplements, have been proposed. Thus, Takeda-Kawaguchi et al recently used a chemically defined medium (MSCGM-CD) to grow isolated human DPSCs that did not have a reduced colony forming ability as compared with that of cells grown in medium supplemented with FBS and had the ability to differentiate into odontoblasts in vitro and to form dentine-like structures when transplanted into immunodeficient mice [ 89 ]. In contrast, some authors have proposed that, pending the availability of completely serum-free defined medium of GMP (“Good Manufacturing Practices”) grade, products derived from secure human blood, such as platelet lysates, might be considered a viable alternative to FBS [ 90 ].…”

Biomedical Application of Dental Tissue‐Derived Induced Pluripotent Stem Cells

“…Exact composition of bovine serum is unknown and varies from batch to batch, resulting in interfering with the reproducibility of iPSCs generation. Moreover, serum could be contaminated with viruses, mycoplasma, prions, or other pathogenic, toxic, or immunogenic agents [ 52 ]. Although little is known regarding other xenogeneic products, porcine-derived trypsin is likely to contain similar biosafety risks.…”