Isolation and Differentiation of Mesenchymal Stem Cells From Broiler Chicken Compact Bones

Abstract

Chicken mesenchymal stem cells (MSCs) can be used as an avian culture model to better understand osteogenic, adipogenic, and myogenic pathways and to identify unique bioactive nutrients and molecules which can promote or inhibit these pathways. MSCs could also be used as a model to study various developmental, physiological, and therapeutic processes in avian and other species. MSCs are multipotent stem cells that are capable of differentiation into bone, muscle, fat, and closely related lineages and express unique and specific cell surface markers. MSCs have been isolated from numerous sources including human, mouse, rabbit, and chicken with potential clinical and agricultural applications. MSCs from chicken compact bones have not been isolated and characterized yet. In this study, MSCs were isolated from compact bones of the femur and tibia of day-old male broiler chicks to investigate the biological characteristics of the isolated cells. Isolated cells took 8–10 days to expand, demonstrated a monolayer growth pattern and were plastic adherent. Putative MSCs were spindle-shaped with elongated ends and showed rapid proliferation. MSCs demonstrated osteoblastic, adipocytic, and myogenic differentiation when induced with specific differentiation media. Cell surface markers for MSCs such as CD90, CD105, CD73, CD44 were detected positive and CD31, CD34, and CD45 cells were detected negative by PCR assay. The results suggest that MSCs isolated from broiler compact bones (cBMSCs) possess similar biological characteristics as MSCs isolated from other chicken tissue sources.

Human fetal skin-derived stem cell secretome enhances radiation-induced skin injury therapeutic effects by promoting angiogenesis

“…In the present study, we selected the stem cells derived from human fetal tissue, which have greater clinical therapeutic potential. CD73, CD90, and CD105 are common cell surface markers of mesenchymal stem cells [32], while Oct4 and Sox2 are the classical markers of embryonic stem cells [33]. In this study, positive expressions of CD73, CD90, CD105, Oct4, and Sox2 were observed in hFSSC which indicated that hFSSC presented with the characteristics of both MSC and embryonic stem cell (ESC).…”

Role of 25-Hydroxyvitamin D3 and 1,25-Dihydroxyvitamin D3 in Chicken Embryo Osteogenesis, Adipogenesis, Myogenesis, and Vitamin D3 Metabolism

“…The samples were analyzed in duplicate by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) performed on an Applied Biosystems StepOnePlus™ (Thermo Fisher Scientific, Waltham, MA, United States) with iTaq™ Universal SYBR Green Supermix (BioRad, Hercules, CA, United States) using the following conditions for all genes: 95°C for 10min followed by 40cycles of 15s denaturation at 95°C, annealing for 20s, and 15s extension at 72°C, followed by 95°C for 15s and a melt curve stage. The key osteogenesis marker genes ( RUNX2 : runt-related transcription factor 2; BMP2 : bone morphogenetic protein 2; COL1A2 : collagen type I alpha 2 chain; BGLAP : bone gamma-carboxyglutamate protein; SPP1 : secreted phosphoprotein 1; and ALP : alkaline phosphatese; Chatakun et al, 2014 ; Adhikari et al, 2019 ); key adipogenesis marker genes ( PPAR-γ : peroxisome proliferator-activated receptor gamma; FASN : fatty acid synthase; and FAPB4 : fatty acid binding protein 4; Bhat et al, 2014 ; Ji et al, 2015 ); key myogenesis marker genes ( MYOG : myogenin; MYOD1 : myogenic differentiation 1, and MYF5 : myogenic factor 5; Wagatsuma and Sakuma, 2014 ); and a vitamin D catabolism gene ( CYP24A1 : cytochrome P450 family 24 subfamily A member 1; Christakos et al, 2010 ) were investigated ( Table 1 ). GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as a housekeeping gene.…”

Mesenchymal Stromal Cells Promote Retinal Vascular Repair by Modulating Sema3E and IL-17A in a Model of Ischemic Retinopathy

“…MSCs characterized by their potential therapeutic properties, easy isolation, wide expansion in vitro , and their abundance in compact bone of many species ( Adhikari et al, 2018 ; Yusop et al, 2018 ) were isolated from the compact bone of mice as previously outlined ( Zhu et al, 2010 ). Long bones of adult C57BL6/J mice (6–8 weeks) were removed and thoroughly cleaned from any connective tissue.…”