In vivo tracking on longer retention of transplanted myocardin gene-modified adipose-derived stem cells to improve erectile dysfunction in diabetic rats

Abstract

Background Stem cell therapy has revealed a promising future for treating erectile dysfunction (ED), but the fate and curative mechanism of intracavernosal transplanted stem cells are under further exploration. This study aimed to demonstrate the effects of myocardin gene modification on improving erectile function and prolonging the retention of implanted adipose-derived stem cells (ASCs) using in vivo small animal imaging. Methods ASCs were isolated, cultured, and identified by flow cytometry and osteogenic and adipogenic induction. The effects of gene modification on cell proliferation, apoptosis, and contraction were determined by CCK-8, EdU, flow cytometry, and collagen gel lattice contraction assays as well as confocal microscopy. A total of 20 normal and 60 diabetes mellitus ED to (DMED) Sprague–Dawley rats were recruited to the 7 day and 21 day groups. Each group contained subgroups of 10 rats each: the negative control (NC), DMED + ASCs plus Ad-Luc-Myocardin, DMED + ASCs plus Ad-Luc, and DMED + phosphate buffer solution (PBS) groups. Erectile function was evaluated with the intracavernosal pressure/mean arterial pressure (△ICP/MAP) ratio. In vivo small animal imaging and an EdU cell tracking strategy were introduced to detect the transplanted ASCs, and IHC and WB were performed to assess smooth muscle cell protein levels. Results The ASCs expressed high CD29 and CD90 and scant CD45, while the multi-induction potential was verified by oil red O and alizarin red staining. Gene transfection of myocardin had no significant influence on ASC apoptosis but inhibited cell proliferation and promoted cell contraction. Myocardin combined with ASCs enhanced the therapeutic potential of ASCs for improving the △ICP/MAP ratio as well as α-SMA and calponin expression. In vivo imaging confirmed that ASCs resided within the cavernous body in 21 days, while only a few red EdU dots were detected. Conclusions Myocardin induced ASC differentiation towards smooth muscle-like cells and enhanced the therapeutic potential of ASCs for ameliorating ED in STZ-induced diabetic rats. Notably, in vivo small animal tracking was an effective strategy for monitoring the implanted stem cells, and this strategy might have advantages over traditional EdU assays. Electronic supplementary material The online version of this article (10.1186/s13287-019-1325-7) contains supplementary material, which is available to authorized users.

Stem cell therapy and diabetic erectile dysfunction: A critical review

“…However, the number of cells injected among these studies ranged from 2 × 10 6 to 5 × 10 5 cells per animal[ 46 , 47 ]. Moreover, in addition to stem cell therapy, several studies added growth factors, hormones and drugs, including insulin[ 48 ], magnetic iron oxide nanoparticles[ 49 ], microtissue[ 50 ], inducible NOS[ 47 ], myocardin[ 51 ], icariin[ 52 ], corin[ 53 ], exosomes from corpus cavernosum smooth muscle cells[ 54 ], fibroblast growth factor 2 (FGF2)[ 55 ], stromal cell-derived cell factor-1 (SDF-1)[ 56 ], VEGF[ 57 ] and hepatocyte growth factor[ 45 ].…”

Antiobesity Effects of Adipose‐Derived Stromal/Stem Cells in a Naturally Aged Mouse Model

“…However, there was no obvious difference in cell proliferation and self‐renewal between the groups. The transplanted ASCs resided within the cavernous body by in vivo tracking (34). These results indicate that transplanted ASCs have antiobesity and antiaging effects by regulating intrinsic stem cell differentiation plasticity and by secreting anti‐inflammatory factors, antioxidant markers, and hormones.…”

NLRP3 downregulation enhances engraftment and functionality of adipose-derived stem cells to alleviate erectile dysfunction in diabetic rats

“…Primary ASCs were obtained as described previously (27). Fat tissue of Sprague-Dawley (SD) rats weighing 180-220 g was isolated from the inguinal area, minced in digestive solution containing 0.15% type I collagenase, and incubated under shaking at 120 rpm in a constant-temperature hybridization oven (UVP, Upland, CA, USA) at, 37°C for 1 h. Subsequently, adipose tissue was resuspended in 10% FBS (HyClone, GE, Boston, MA,USA) complete medium to terminate the digestion and washed with PBS by centrifugation at 1200 rpm, then processed with FACS Lysing Solution (BDIS Catalog No.349202) for 10 min.…”

“…Male SD rats (approximately 200 g and 8-weeks-old) with normal erectile function were provided by the Experimental Animal Center of Southern Medical University. The rats with DMED were modeled and identified as previously described (27). Briefly, 80 healthy adult male SD rats were intraperitoneally injected with 1% streptozotocin solution (65 mg kg −1 ).…”