TY - JOUR
T1 - Chondrogenic and Osteogenic In Vitro Differentiation Performance of Unsorted and Sorted CD34+, CD146+, and CD271+ Stem Cells Derived from Microfragmented Adipose Tissue of Patients with Knee Osteoarthritis
AU - Bagge, Jasmin
AU - Mahmood, Haider
AU - Janes, Jennifer
AU - Vomstein, Kilian
AU - Blønd, Lars
AU - Hölmich, Lisbet R
AU - Freude, Kristine
AU - Nehlin, Jan O
AU - Barfod, Kristoffer W
AU - Hölmich, Per
PY - 2025/2/11
Y1 - 2025/2/11
N2 - Background/Objectives: Treatment of knee osteoarthritis (OA) with autologous stem cells from microfragmented adipose tissue (MFAT) has shown promising but varying results. Multiple stem cell types, including CD34+, CD146+, and CD271+ stem cells, have been identified within MFAT. Patient-specific heterogeneity in stem cell populations and the content of highly potent cells may be determining factors for a successful treatment outcome. The current study aimed to identify the most promising stem cell type in MFAT to treat OA, focusing on their chondrogenic and osteogenic differentiation performance. Methods: CD34+, CD146+, and CD271+ stem cells from the MFAT of eight patients with knee OA were separated using magnetic-activated cell sorting (MACS) and analyzed as subtypes. Unsorted cells were used as a control. Chondrogenic and osteogenic in vitro differentiation were assessed through Safranin-O and H&E staining, pellet size, and qPCR for chondrogenesis, as well as Alizarin Red S staining and qPCR for osteogenesis. Results: CD34+, CD146+, and CD271+ stem cells were doubled using MACS. All subtypes were able to undergo osteogenic differentiation with Alizarin Red S staining, revealing a significant increase in calcium deposits of induced cells compared to non-induced controls. CD146+ stem cells showed higher calcium deposition compared to CD34+, CD271+, and unsorted stem cells. All cell types could form chondrogenic pellets. CD271+ stem cells produced more proteoglycans, as shown by Safranin-O staining, than CD34+ and CD146+ stem cells, but not more than the unsorted stem cells. After differentiation induction, all cell types showed an upregulation of most chondrogenic and osteogenic biomarkers. Conclusions: CD146+ stem cells showed the highest osteogenic differentiation performance for calcium deposition, while CD271+ stem cells showed the greatest chondrogenic differentiation performance for proteoglycan formation. The prevalence of these stem cell types may play a critical role in the clinical effectiveness when treating OA.
AB - Background/Objectives: Treatment of knee osteoarthritis (OA) with autologous stem cells from microfragmented adipose tissue (MFAT) has shown promising but varying results. Multiple stem cell types, including CD34+, CD146+, and CD271+ stem cells, have been identified within MFAT. Patient-specific heterogeneity in stem cell populations and the content of highly potent cells may be determining factors for a successful treatment outcome. The current study aimed to identify the most promising stem cell type in MFAT to treat OA, focusing on their chondrogenic and osteogenic differentiation performance. Methods: CD34+, CD146+, and CD271+ stem cells from the MFAT of eight patients with knee OA were separated using magnetic-activated cell sorting (MACS) and analyzed as subtypes. Unsorted cells were used as a control. Chondrogenic and osteogenic in vitro differentiation were assessed through Safranin-O and H&E staining, pellet size, and qPCR for chondrogenesis, as well as Alizarin Red S staining and qPCR for osteogenesis. Results: CD34+, CD146+, and CD271+ stem cells were doubled using MACS. All subtypes were able to undergo osteogenic differentiation with Alizarin Red S staining, revealing a significant increase in calcium deposits of induced cells compared to non-induced controls. CD146+ stem cells showed higher calcium deposition compared to CD34+, CD271+, and unsorted stem cells. All cell types could form chondrogenic pellets. CD271+ stem cells produced more proteoglycans, as shown by Safranin-O staining, than CD34+ and CD146+ stem cells, but not more than the unsorted stem cells. After differentiation induction, all cell types showed an upregulation of most chondrogenic and osteogenic biomarkers. Conclusions: CD146+ stem cells showed the highest osteogenic differentiation performance for calcium deposition, while CD271+ stem cells showed the greatest chondrogenic differentiation performance for proteoglycan formation. The prevalence of these stem cell types may play a critical role in the clinical effectiveness when treating OA.
KW - CD146 stem cells
KW - CD271 stem cells
KW - CD34 stem cells
KW - differentiation potential
KW - magnetic-activated cell sorting
KW - microfragmented adipose tissue
KW - osteoarthritis
KW - pericytes
UR - http://www.scopus.com/inward/record.url?scp=85219166083&partnerID=8YFLogxK
U2 - 10.3390/jcm14041184
DO - 10.3390/jcm14041184
M3 - Journal article
C2 - 40004714
SN - 2077-0383
VL - 14
JO - Journal of Clinical Medicine
JF - Journal of Clinical Medicine
IS - 4
M1 - 1184
ER -