Successful isolation of viable stem cells from cryopreserved microfragmented human adipose tissue from patients with knee osteoarthritis - a comparative study of isolation by tissue explant culture and enzymatic digestion.

Autor: Bagge J; Sports Orthopedic Research Center - Copenhagen (SORC-C), Department of Orthopedic Surgery, Copenhagen University Hospital - Amager and Hvidovre, Kettegård Allé 30, 2650, Hvidovre, Denmark. jasmin.bagge@regionh.dk., Hölmich P; Sports Orthopedic Research Center - Copenhagen (SORC-C), Department of Orthopedic Surgery, Copenhagen University Hospital - Amager and Hvidovre, Kettegård Allé 30, 2650, Hvidovre, Denmark., Hammer FA; Sports Orthopedic Research Center - Copenhagen (SORC-C), Department of Orthopedic Surgery, Copenhagen University Hospital - Amager and Hvidovre, Kettegård Allé 30, 2650, Hvidovre, Denmark., Nehlin JO; Department of Clinical Research, Copenhagen University Hospital - Amager and Hvidovre, Kettegård Allé 30, 2650, Hvidovre, Denmark., Vomstein K; Department of Obstetrics and Gynecology, The Fertility Clinic, Copenhagen University Hospital - Hvidovre, Kettegård Allé 30, 2650, Hvidovre, Denmark., Blønd L; Department of Orthopedic Surgery, Zealand University Hospital - Køge, Lykkebækvej 1, 4600, Køge, Denmark., Hölmich LR; Department of Plastic Surgery, Copenhagen University Hospital - Herlev and Gentofte, Borgmester Ib Juuls Vej 1, 2730, Herlev, Denmark., Barfod KW; Sports Orthopedic Research Center - Copenhagen (SORC-C), Department of Orthopedic Surgery, Copenhagen University Hospital - Amager and Hvidovre, Kettegård Allé 30, 2650, Hvidovre, Denmark.
Jazyk: angličtina
Zdroj: Journal of experimental orthopaedics [J Exp Orthop] 2023 Mar 23; Vol. 10 (1), pp. 31. Date of Electronic Publication: 2023 Mar 23.
DOI: 10.1186/s40634-023-00596-x
Abstrakt: Purpose: To investigate if viable stem cells could be isolated and expanded from cryopreserved microfragmented adipose tissue (AT) harvested from patients with knee osteoarthritis.
Methods: Microfragmented abdominal AT from knee osteoarthritis patients was cryopreserved at -80 °C in cryoprotectant-medium. The samples were thawed for stem cell isolation by tissue explant culture (TEC) and enzymatic digestion (ED), respectively. Viability, population doublings, and doubling time were assessed by trypan blue staining and flow cytometry. Cell type and senescence-associated β-galactosidase activity were analyzed by flow cytometry. Osteogenic and adipogenic differentiation was assessed quantitatively by Alizarin-Red-S and Oil-Red-O staining, respectively.
Results: Microfragmented AT from 7 patients was cryopreserved for a period of 46-150 days (mean (SD) 115.9 days (44.3 days)). Viable stem cells were successfully recovered and expanded from all patients using both isolation methods with no significant difference in viable population doublings or doubling time from passage 1 to 3 (p > 0.05). Low levels of senescence-associated β-galactosidase activity was detected for both methods with no significant difference between TEC and ED (p = 0.17). Stemness was verified by stem cell surface markers and osteogenic and adipogenic differentiation performance. Adventitial stem cells (CD31 - CD34 + CD45 - CD90 + CD146 - ), pericytes (CD31 - CD34 - CD45 - CD90 + CD146 + ), transitional pericytes (CD31 - CD34 + CD45 - CD90 + CD146 + ), and CD271 + stem cells (CD31 - CD45 - CD90 + CD271 + ) were identified using both methods. More pericytes were present when using TEC (25% (24%)) compared to ED (3% (2%)) at passage 4 (p = 0.04).
Conclusions: Viable stem cells can be isolated and expanded from cryopreserved microfragmented AT using both TEC and ED. TEC provides more clinically relevant pericytes than ED.
(© 2023. The Author(s).)
Databáze: MEDLINE
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