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Journal of Experimental Orthopaedics
Fig. 5 | Journal of Experimental Orthopaedics

Fig. 5

From: 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

Fig. 5

Cellular senescence evaluated by senescence associated-β-galactosidase (SA-BGAL) activity. SA-BGAL activity of cryopreserved microfragmented adipose tissue (AT) when analyzed quantitatively using flow cytometry at passage 4. (A) Forward (FSC-A)- and side-scatter (SSC-A) plot of senescent positive control sample (see Methods). P1 gating was used to remove debris and gate on non-P1 cells. Green colored cells (marked by ellipse) show backgating of identified senescent cells of large size and high granularity. (B) FITC histogram of unstained and stained positive control senescent cells used for SA-BGAL activity gating. (C) Box plot of the quantity (%) of cells with SA-BGAL activity from cryopreserved microfragmented AT when isolated with tissue explant culture (TEC) or enzymatic digestion (ED). ns: non-significant when analyzed with a paired t-test. (D) Histogram showing gating of SA-BGAL + cells as a measure of senescence from the senescence positive control. (E) Representative histogram showing SA-BGAL + cells as a measure of cellular senescence from cryopreserved microfragmented AT when isolated by TEC, and (F) when isolated by ED

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