Isolation and expansion of synovial CD34(-)CD44(+)CD90(+) mesenchymal stem cells: comparison of an enzymatic method and a direct explant technique.
Synovium-derived mesenchymal stem cells (MSCs) offer a promising therapeutic option for cartilage regeneration. The conventional method of MSC isolation involves single-cell suspensions using collagenases. Recently, a nonenzymatic explant technique was developed to isolate MSCs. We compared these techniques in the isolation of functional MSCs. MSCs were isolated from human fibrous and adipose synovium of osteoarthritic patients using explants or enzymatic methods. Total cell number, percentage of MSCs, and surface marker expression of MSCs were measured following expansion. Multipotentiality was determined using a MSC functional identification kit. MSCs isolated from fibrous or adipose synovium using these two techniques expressed similar levels of the surface markers CD44, CD90, and CD105, and displayed similar multipotentiality in generating adipocytes, osteoblasts, and chondrocytes. Total cell number and number of CD34(-)CD44(+)CD90(+) MSCs after 10-day expansion were similar in each culture, regardless of the source and method used, although the percentage of MSCs was slightly higher in explant cultures. There were no correlations between MSC yield and patient age, Hospital for Special Surgery score, and degree of deformity under all culture conditions. Both the enzymatic and explant techniques yielded similar yields of MSCs with similar characteristics. Because the explant technique is simpler and less invasive, it may be preferred over enzymatic techniques for isolating MSCs from the synovium of osteoarthritic patients for cartilage regeneration.
Department of Orthopaedic Surgery, Korea University College of Medicine , Seoul , Korea.
Dae-Hee
DH
Lee
Seong-Dae
SD
Joo
Seung-Beom
SB
Han
Jisu
J
Im
Soon-Hyuck
SH
Lee
Chung Hee
CH
Sonn
Kyung-Mi
KM
Lee
D000273:Adipose Tissue / Q000166:cytology; D000368:Aged; D000369:Aged, 80 and over; D015703:Antigens, CD / Q000378:metabolism*; D018952:Antigens, CD34 / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D017364:Collagenases / Q000378:metabolism*; D003710:Demography; D005260:Female; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008875:Middle Aged; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D013582:Synovial Fluid / Q000166:cytology; D018800:Thy-1 Antigens / Q000378:metabolism
D015703:Antigens, CD; D018952:Antigens, CD34; D018960:Hyaluronan Receptors; D018800:Thy-1 Antigens; D017364:Collagenases