Int J Stem Cells.  2023 Feb;16(1):66-77. 10.15283/ijsc21146.

Efficacy and Safety of Human Bone Marrow-Derived Mesenchymal Stem Cells according to Injection Route and Dose in a Chronic Kidney Disease Rat Model

• Chae HK
• Suh N
• Jang MJ
• Kim YS
• Kim BH
• Aum J
• Shin HC
• You D
• Hong B
• Park HK
• Kim CS

• KMID: 2539622
• DOI: http://doi.org/10.15283/ijsc21146

Abstract

Background and Objectives
We compared the efficacy and safety of human bone marrow-derived mesenchymal stem cells (hBMSC), delivered at different doses and via different injection routes in an animal model of chronic kidney disease.
Methods and Results
A total of ninety 12-week-old rats underwent 5/6 nephrectomy and randomized among nine groups: sham, renal artery control (RA-C), tail vein control (TV-C), renal artery low dose (RA-LD) (0.5×10 ⁶ cells), renal artery moderate dose (RA-MD) (1.0×10 ⁶ cells), renal artery high dose (RA-HD) (2.0×10 ⁶ cells), tail vein low dose (TV-LD) (0.5×10 ⁶ cells), tail vein moderate dose (TV-MD) (1.0×10 ⁶ cells), and tail vein high dose (TV-HD) (2.0×10 ⁶ cells). Renal function and mortality of rats were evaluated after hBMSC injection. Serum blood urea nitrogen was significantly lower in the TV-HD group at 2 weeks (p＜0.01), 16 weeks (p＜0.05), and 24 weeks (p＜0.01) than in the TV-C group, as determined by one-way ANOVA. Serum creatinine was significantly lower in the TV-HD group at 24 weeks (p＜0.05). At 8 weeks, creatinine clearance was significantly higher in the TV-MD and TV-HD groups (p＜0.01, p＜0.05) than in the TV-C group. In the safety evaluation, we observed no significant difference among the groups.
Conclusions
Our findings confirm the efficacy and safety of high dose (2×10 ⁶ cells) injection of hBMSC via the tail vein.

Keyword

Chronic kidney disease; Stem cell transplantation; Cell migration; Bone marrow

Figure

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  Fig. 1 Flow cytometric histograms, morphology of hBMSC and efficacy and safety evaluation schedule. (A) FACS data of hBMSC. (B) Morphology of hBMSC and osteogenic, adipogenic differentiation lineage. (C) Experimental schedule. RA: renal artery, RA-C: renal artery control, RA-LD: renal artery low dose, RA-MD: renal artery moderate dose, RA-HD: renal artery high dose, TV-C: tail vein control, TV-LD: tail vein low dose, TV-MD: tail vein moderate dose, TV-HD: tail vein high dose, PSA: plasma solution A, hBMSC: human bone marrow-derived mesenchymal stem cells.

• 

  Fig. 2 Efficacy of hBMSC in CKD rat model according to injection route and hBMSC dose. BUN: blood urea nitrogen, Ccr: Creatinine clearance, RA-C: renal artery control, RA-LD: renal artery low dose, RA-MD: renal artery moderate dose, RA-HD: renal artery high dose, TV-C: tail vein control, TV-LD: tail vein low dose, TV-MD: tail vein moderate dose, TV-HD: tail vein high dose. *p<0.05; **p<0.01; ***p<0.001, vs. the RA-C group; *p<0.05; **p<0.01; ***p<0.001, vs. the TV-C group; Red colored stars indicate statistically significant differences in RA-LD, RA-MD, RA-HD, groups compared to the RA-C group, respectively. Blue colored stars indicate statistically significant differences in TV-LD, TV-MD, TV-HD, groups compared to the TV-C group, respectively. (A∼H) 3 times per rat, 10 rats per group, but 9 rats in the RA-C, TV-LD, TV-HD groups and 8 rats in the TV-MD groups.

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  Fig. 3 Histopathological analysis of the renal artery injection group. (A) Histopathological scoring, cortex. (B) Histopathological scoring, medulla. (C) Sirius red, cortex. (D) Sirius red, medulla. (E) TUNEL, cortex. (F) TUNEL, medulla. (G) α-SMA, cortex. (H) α-SMA, medulla. (I) ED-1, cortex. (J) ED-1, medulla. Non-overlapping fields (400×). *p<0.05; **p<0.01; ***p<0.001, vs. the RA-C group; $$p<0.01, vs. the RA-LD group; RA-C: renal artery control, RA-LD: renal artery low dose, RA-MD: renal artery moderate dose, RA-HD: renal artery high dose. (A, B) 10 fields per rat, 10 rats per group, but 9 rats in the RA-C, TV-LD, TV-HD groups and 8 rats in the TV-MD groups. (C∼J) 4 fields per rat, 10 rats per group, but 9 rats in the RA-C, TV-LD, TV-HD groups and 8 rats in the TV-MD groups.

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  Fig. 4 Histopathological analysis of the tail vein injection group. (A) Histopathological scoring, cortex. (B) Histopathological scoring, medulla. (C) Sirius red, cortex. (D) Sirius red, medulla. (E) TUNEL, cortex. (F) TUNEL, medulla. (G) α-SMA, cortex. (H) α-SMA, medulla. (I) ED-1, cortex. (J) ED-1, medulla. Non-overlapping fields (400×). NS, p>0.05; *p<0.05; **p<0.01; ***p<0.001 vs. the TV-C group; $$p<0.01, vs. the TV-LD group; TV-C: tail vein control, TV-LD: tail vein low dose, TV-MD: tail vein moderate dose, TV-HD: tail vein high dose. (A, B) 10 fields per rat, 10 rats per group, but 9 rats in the RA-C, TV-LD, TV-HD groups and 8 rats in the TV-MD groups. (C∼J) 4 fields per rat, 10 rats per group, but 9 rats in the RA-C, TV-LD, TV-HD groups and 8 rats in the TV-MD groups.

Reference

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