Isolation and characterization of mesenchymal stem cells in orthopaedics and the emergence of compact bone mesenchymal stem cells as a promising surgical adjunct

Table 1 Source and isolation methods of mesenchymal stromal cells

Source	Isolation method	Ref.
Bone marrow	Aspirates cultured and media changed every 3-4 d to select for MSCs	[34]
	Aspirates layered over Ficoll-Paque density-gradient and plates in tissue culture dish. Adherent cells maintained with periodic passaging	[82]
	Bone marrow mononuclear cells seeded from single colony-forming unit fibroblasts and selected for by CD105(+)/CD45(-)	[83]
	Sort bone mononuclear cells based on aldehyde deydrogenase expression (ALDHhighCD45-)	[84]
	Sort based on CD45−/lowCD271+ phenotype following a microbead-based pre-enrichement	[85]
	Layer bone marrow over hyluronic acid followed by centrifugation, collect most superficial layer containing the mononuclear cells	[86]
Compact bone	Trabecular bone fragments rinsed and placed in complete α-MEM/Ham’s F12, confluent monolayers were obtained within 10-20 d	[87-89]
	Bone cell cultures established by treating bone fragments with collagenase in low Ca2+ medium	[9,90,30]
	Compact bone fragments obtained, cultured, and isolated. CB-MSCs then undergo trypsinization to reveal enhanced osteogenic capacity	[43]
Adipose	Place 10-20 mL of washed adipose tissue in 100 mm Petri dish; dissect out yellow tissue; mince tissue finely and place in enzymatic digestion solution; centrifuge and collect pellet for wash; resuspend in complete culture medium	[14,13]
	Wash lipoaspirate with PBS; enzymatically digest using collagenase 1A solution; spin down cells, wash and plate in complete medium	[12]
Bone marrow	Aspirates cultured and media changed every 3-4 d to select for MSCs	[34]
	Aspirates layered over Ficoll-Paque density-gradient and plates in tissue culture dish. Adherent cells maintained with periodic passaging	[82]
	Bone marrow mononuclear cells seeded from single colony-forming unit fibroblasts and selected for by CD105(+)/CD45(-)	[83]
	Sort bone mononuclear cells based on aldehyde deydrogenase expression (ALDHhighCD45-)	[84]
	Sort based on CD45−/lowCD271+ phenotype following a microbead-based pre-enrichement	[85]

MSC: Mesenchymal stromal cell; CB: Compact bone; PBS: Phosphate buffer saline.