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

Jasmin Bagge*, Haider Mahmood, Jennifer Janes, Kilian Vomstein, Lars Blønd, Lisbet R Hölmich, Kristine Freude, Jan O Nehlin, Kristoffer W Barfod, Per Hölmich

*Corresponding author for this work

Abstract

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.

Original languageEnglish
Article number1184
JournalJournal of Clinical Medicine
Volume14
Issue number4
ISSN2077-0383
DOIs
Publication statusPublished - 11 Feb 2025

Keywords

  • CD146 stem cells
  • CD271 stem cells
  • CD34 stem cells
  • differentiation potential
  • magnetic-activated cell sorting
  • microfragmented adipose tissue
  • osteoarthritis
  • pericytes

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