Gingival-Derived Mesenchymal Stem Cell from Rabbit (Oryctolagus cuniculus): Isolation, Culture, and Characterization

Abstract

Objective This study aims to confirm whether the GDMSCs isolated from rabbit’s (Oryctolagus cuniculus) gingiva are mesenchymal stem cells (MSCs). Materials and Methods This study design was partly quasi-experimental with an observational design. GDMSCs were isolated from the gingiva of healthy male rabbits (O. cuniculus) (n = 2), 6 months old, and 3 to 5 kg of body weight. The specific cell surface markers of MSCs; clusters of differentiation (CD), namely, CD44, CD73, CD90, CD105, and CD200 expressions; and hematopoietic stem cell surface markers CD34 and CD45 were examined using flow cytometry and immunohistochemistry with immunofluorescence. The osteogenic differentiation of isolated GDMSCs was examined using alizarin red staining. Results GDMSCs in the fourth passage showed a spindle-like formation and fibroblast-like cells that attached to the base of the culture plate. GDMSCs were MSCs that positively expressed CD44, CD73, CD90, CD105, and CD200 but did not express CD34 and CD45 when examined using flow cytometry and immunohistochemical analysis. GDMSCs had osteogenic differentiation confirmed by calcified deposits in vitro with a red–violet and brownish color after alizarin red staining. Conclusion GDMSCs isolated from the rabbits (O. cuniculus) were confirmed as MSCs in vitro documented using immunohistochemistry and flow cytometry. GDMSCs can differentiate into osteogenic lineage in vitro that may be suitable for regenerative dentistry.

Comparison of biological features of wild European rabbit mesenchymal stem cells derived from different tissues

“…Considering this, we have selected a panel of antibodies that have shown high sensitivity and repeatability and confirmed these results through RT-PCR assessment of expression at the mRNA level. Our data demonstrate that all rMSCs isolated were negative for CD34 and CD45 but positive for CD44, CD29, NANOG, OCT4, and SOX2, coinciding with the results reported in the literature for rMSCs isolated from different tissues: adipose [19], gingival [20], amniotic fluid [15] and bone marrow [21,23]. The basic in vitro trilineage differentiation capacity of rMSC, that is adipocytes, osteocytes, and chondrocytes have been reported in the literature only for adipose [18] and for bone marrow rMSC [23][24][25].…”

“…Postnatal organs and tissues are good MSC sources; however, each source of MSCs has a different degree of differentiation potential and expression of a different set of stem cell-related markers, as well as other important characteristics such as high proliferation, immunomodulation, and allo-and xeno-transplantation ability. Rabbit MSCs (rMSC) have been widely used as preclinical models for orthopedic, specifically bone, articular cartilage, ligament reconstruction, and spinal fusion, as well as cardiovascular regenerative medicine strategies [18][19][20][21]. Although rMSCs are being investigated in these models, they have not been fully characterized in terms of immunophenotype and differentiation potential, and many tissues have not yet been explored as a source of MSC in the rabbit.…”

“…MSCs have been found in some rabbit tissues (adipose, gingival, bone marrow, Wharton's jelly, amniotic fluid) exhibiting stem cell characteristics such as plastic adherence, multilineage differentiation capacity, MSC marker expression, and pluripotent gene expression [20][21][22][23]. Postnatal organs and tissues are good MSC sources; however, each source of MSCs has a different degree of differentiation potential and expression of a different set of stem cell-related markers, as well as other important characteristics such as high proliferation, immunomodulation, and allo-and xeno-transplantation ability.…”

Clinical Applications of Cell-Scaffold Constructs for Bone Regeneration Therapy

“…Hence, the characterization of MSCs based on surface marker analysis is an essential criterion for the clinical application of BTE methodologies. According to the International Society of Cell Therapy (ISCT) criteria, MSCs express a cluster of differentiation (CD) surface markers such as CD90, CD105, and CD73, but do not express CD11b, CD14, CD19, CD34, CD45, or human leukocyte antigen (HLA)-DR [32][33][34]. However, this set of cell surface markers is not always useable for the identification of MSCs.…”

Elemental and Configural Associative Learning in Spatial Tasks: Could Zebrafish be Used to Advance Our Knowledge?

“…This type of learning may have high fitness value in rapidly changing, spatially complex, i.e., natural, environments. Spatial learning has been demonstrated in multiple contexts including migration (Winkler et al, 2014;Lindecke et al, 2019;Franzke et al, 2020), foraging (Croney et al, 2003;D'Adamo and Lozada, 2003;Buatois and Lihoreau, 2016), territoriality and reproductive behavior (Füller et al, 1983;Hardenberg et al, 2000). Not surprisingly, spatial learning is found in a variety of species from insects (Wystrach, 2018) to humans (Keller and Just, 2016;Piber et al, 2018).…”