Prospective Isolation of a Functional Human Cartilage Progenitor Capable of Forming Hyaline Cartilage In Vivo
Lauren Koepke, B.S. Cellular/Molecular Biology.
Stanford University, San Jose, CA, USA.
Purpose: Over the past century, the average life span has increased by nearly 30%, leading to an overall aging population and an increase in degenerative diseases, including osteoarthritis. Due to a lack of vasculature and low cellularity in articular cartilage, regeneration does not occur. Since we have identified three human chondroprogenitor (CP) populations, we have the ability to functionally characterize these CPs to identify if these populations give rise to unique types of cartilage.
Methods: Prospective FACS analysis was performed on digested fresh human bone specimens. The gating strategy was verified in vivo and in vitro. The isolated cell gene expression profile of each population was performed using q-PCR and microarray. In vitro differentiation was performed and analyzed via FACS. Cellular proliferation using CFU in vitro and EdU both in vitro and in vivo was performed. In vivo differentiation was performed using our previously published renal capsule model as well as our xenograft model. The in vivo samples were then FACS sorted for analysis and corresponding IHC (col2, col10, mmp13) and histology (safranin-o fast green and pentachrome) was performed.
Results: Three distinct populations were successfully isolated. Each cell population had a unique gene expression profile signifying a possible variance in differentiation capability. In vitro as well as in vivo differentiation varied between each population CP1-3. The relative proliferation variability also suggests inherent population differences.
Conclusion: The potential to isolate a particular CP population that is most proliferative and has the capacity to give rise to articular cartilage is exciting and may provide a new therapeutic strategy for treating patients with osteoarthritis.
Figure 1. FACS gating strategy for our three human chondroprogenitor populations, CP1-3.
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