Long term explant culture for harvesting homogeneous population of human dental pulp stem cells

Abstract

Dental pulp stem cells have emerged as a preferred source of mesenchymal stem cells, because of its easy availability and high stem cell content. Dental pulp is a specific fibrous tissue that contains heterogeneous populations of odontoblasts, fibroblasts, pericytes, progenitors, stem cells, leukocytes and neuronal cells. In this study, we propose sustained explant culture as a simple, economical and efficient process to isolate dental pulp stem cells from human Dental pulp Tissue. Historically explant cultures were used to get fibroblast cells from embryonic chick heart using plasma clot cultures. The subculture was performed by lifting mother explant (original explant) and grafting it in a new plasma clot. We modified this age old technique to suit the modern times. Here we demonstrate for the first time that the mother explant (E0) of human dental pulp tissue could be sub‐cultured consecutively seven times (E7) without displacement. This technique is highly reproducible and permits growth and proliferation of dental pulp stem cells yielding an enriched homogeneous mesenchymal stem cells population in the first passage itself as revealed by surface marker expression. These dental pulp stem cells exhibit differentiation into adipogenic, chondrogenic and osteogenic lineage revealing their mesenchymal stem cell nature. We propose that dental pulp stem cells isolated by sustained explant culture are phenotypically and functionally comparable to those obtained by enzymatic method. It is a simple, inexpensive and gentle method, which may be preferred over the conventional techniques for obtaining stem cells from other tissue sources as well especially in cases of limited starting material.

Effect of Ascorbic Acid on Differentiation, Secretome and Stemness of Stem Cells from Human Exfoliated Deciduous Tooth (SHEDs)

“…Isolation and characterization of SHEDs were carried out using the explant culture method previously described [ 4 ]. Pulp tissue was minced into tiny fragments and placed in 35 mm polystyrene plastic culture dishes.…”

“…Mesenchymal stem cells (MSCs) are pluripotent progenitor cells that undergo mitosis several times and eventually differentiate into osteoblasts, chondrocytes, and adipocytes, etc. MSCs are currently being considered for the management of various diseases [ 1 , 2 , 3 , 4 ]. Moreover, the secretome of mesenchymal stem cells contains several cytokines, growth factors that are responsible for the various effects of transplantation of stem cells.…”

“…They are considered a valuable source for cellular banking and longstanding cryopreservation [ 25 , 26 , 27 ]. The characteristics of SHEDs include greater propagation rate, enhanced clonogenicity, and elevated osteoinductive capacity in vivo in comparison with other types of MSCs [ 4 , 28 ]. Studies have demonstrated that oral MSCs derived from human dental pulp exhibit in vitro pluripotency and can differentiate into several cell populations, including odontoblast, chondrocytes, osteoblasts, and neural cells.…”

Viability of Quercetin-Induced Dental Pulp Stem Cells in Forming Living Cellular Constructs for Soft Tissue Augmentation

“…Recently, a more cost-effective approach to cell sheet formation, by culturing cells under the high-density condition on regular culture dishes, has been propounded and successively utilized [ 4 , 5 , 6 ]. The cell sheet technology has the potential to rectify the conundrum of scaffold fabrication [ 4 , 7 , 8 ]. The biggest setback of scaffold-based tissue engineering is the inflammatory response post-implantation, which hinders the utility of scaffolds and tissue engineering processes [ 9 , 10 ].…”

Tissue engineering: A new era in dentistry

“…Tissue engineering in dentistry has shown remarkable progress, especially due to successful isolation and characterization of stem cells from the oral tissues including dental pulp, gingiva. Dental pulp derived mesenchymal stem cells have shown to undergo tripod differentiation (chondrocyte, osteocyte, adipocyte) 2,3 . Pluripotency of these stem cells has played a major role in the regeneration of wide range of oral tissues.…”