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Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: Investigation of stemness and multipotency of equine adipose-derived mesenchymal stem cells (ASCs) from different fat sources in comparison with lipoma

Fig. 3

Evaluation of stemness of ASCs from various fat sources. CFU assay of subcutaneous (SC), retroperitoneal (RP), and lipoma (LP) adipose-tissue derived ASCs cultivated as 100 cells per 25 cm2 culture flasks for 8 days in growth medium composed of 10% FCS in DMEM and 1% P/S. ac Representative microscopic images of SC, RP, and LP derived colonies stained with 1% crystal violet (blue) show colony formation in all experimental groups. d The number of 1% crystal violet stained colonies were counted. The analysis shows a marked CFU capacity of SC and RP fat-derived cells compared to those cells of LP. ei Evaluation of mesenchymal stem relative markers expression. Subcutaneous (SC), retroperitoneal (RP), and lipoma (LP) adipose tissue-derived ASCs were cultivated under growth conditions for 48 h. A volume of 1 μg RNA per experimental group was transcribed into cDNA using a reverse transcription kit. The expression of CD44 (e), CD90 (f), CD105 (g), CA9 (h), and osteopontin (OPN, i) was quantified using RT-qPCR. The analysis reveals upregulation of the stem cell markers CD44, CD90, and CD105 in RP and SC derived cells compared to LP cells (eg). Upregulation of the CA9 and OPN expression could be shown only in LP derived cells (h, i). The analysis was performed in triplicates for all experimental groups. GAPDH and 18S house-keeping genes were used as endogenous references. The data presented as mean ± SEM. *p < 0.05 and **p < 0.01. Scale bar = 5 mm

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