(Mesenchymal) Stem Cell-Based Therapy in Cisplatin-Induced Acute Kidney Injury Animal Model: Risk of Immunogenicity and Tumorigenicity

Table 1

Xenotransplantation of human stem cells into immunocompromised or immunocompetent cisplatin-treated rodents.

Species, strain	Cisplatin treatment	Stem cell source	Stem cell treatment	Route	hMSC criteria	End	Results: effect of treatment on measured parameters	Cell tracking	Ref.
Sprague-Dawley	D0: 5 mg/kg, ip	hUC-derived exosomes	0.5 h prior cis; 0.2 mg	rsc	No	D3	Blood: ↓ BUN, Cr, TNFα, IL-1β, IL6; kidney: ↓ histology score, TUNEL, ↑ PCNA, Bax, LC3B, BCL-2 (autophagy)	No	[7]
C57BL/6; n = 6-7	D0: 22 mg/kg, sc (20% mortality on D6)	hUCB versus mouse BM (ip); (third party, allogenic)	D1: 1 × 10^6	iv/ip; ip	Yes	D3	Preventive effect regardless of delivery route (xenogenic: iv/ip) or MSC source (xeno/allogeneic: ip); Blood: ↓ BUN, Cr; kidney: ↓ MCP-1, IL-6, TNFα, ↑ IL-10, VEGF, ≈IL-2, ↑ Treg in the kidney and spleen; Mouse BM ≈ MCP-1, IL-6, IL-2, ↓ TNFα, ↑ IL-10, VEGF, ↑ Treg in the kidney and spleen	PKH-26: D3: observed in the kidney and spleen but not in the lung or peritoneum	[8]
C57BL/6; n = 6-7	D0: 20 mg/kg, sc	hUCB versus mouse BM (third party)	D3: 1 × 10^6	iv/ip; ip	Yes	D6	MSC treatment after established renal dysfunction did not show any effect	No	[8]
Sprague-Dawley	D0: 5 mg/kg, ip	Rat BM; hAd; hAF	D1: 5 × 10^6 in 0.5 ml CM	iv	No	D4^p; D11; D30	Comparable therapeutic effects of allogenic and xenogeneic MSCs; Blood: ↓ BUN, Cr; kidney: ↓ histology score, ↓ MDA, ↑ GSH, SOD; urine: ↑ Cr clearance	No	[9]
Sprague-Dawley	D0: 6 mg/kg, ip	hAd	D1: 1-2 × 10^6 in 1 ml saline	iv	No	D5	Blood: ↓ BUN, Cr; kidney: ↓ histology score, ↓ TUNEL, ↑ PCNA; Urine: ↓ mALB, β2 mG	PKH-26 and CD105: rare around kidney tubules, frequent in the liver and spleen	[10]
C3H, female; 25–30 g, n = 5–7	D0: 15 mg/kg, ip	hBM alone or with pFUS	D1: 1 × 10^6	iv	No	D4	MSCs alone had weak positive effect (blood: ↓ BUN, ≈Cr; kidney: ≈histology score, ≈pAKT, ≈Ki-67, ↓ TUNEL); MSCs + pFUS had improved effect (blood: ↓ BUN, Cr; kidney: ↓ histology score, TUNEL, pAKT, Ki-67)	Human mitochondria^+ cells: 24 h: peritubular space (4–10 MSC/field); D3: 2–4/field	[11]
C3H, female; 25–30 g, n = 5–7	D0: 15 mg/kg, ip	hBM; (pFUS)	D3: 1 × 10^6	iv	No	D7	MSCs alone had no significant effect; MSCs + pFUS had improved effect: ↑ survival; blood: ↓ dBUN, Cr	Human mitochondria^+ cells: 24 h: peritubular space (2-fold higher number detected)	[11]
White albino rats	D0: 5 mg/kg, ip	hUC-derived hematopoietic stem cells (CD34^+)	D1: 3 × 10^6	ip	No	2 wk; 4 wk	Blood: ↓ BUN, Cr, K, Na; kidney: ↑ HGF, IGF-1, VEGF, P53	PKH-26: detected in kidney; Time: not stated	[12]
Sprague-Dawley	D0: 6 mg/kg, ip	hUCB	D1: 2 × 10^6 in 0.5 ml saline	iv	No	D5; 6 wk; 8 wk	Blood: ↓ BUN, Cr; kidney: ↑ PCNA, Bcl-2, ↓ TUNEL, Bax, IL-1β, TNFα, MDA, ↓ histology score,; D10: both groups returned to baseline level (BUN, Cr); 6 wk and 8 wk: structural restoration better after MSCs, increased Bax/Bcl-2 ration, ↓ TGF-β1, Masson, ↓ vimentin, ↑ E-cadherin	CM-Dil (cross-linkable membrane dye) in vivo imaging of labeled cells in kidney; Time: 4 wks	[13]
C57BL/6J	D0: 10 mg/kg ip; D1: 10 mg/kg ip	Mesenchymal-like progenitor cells from hESC	D3: 5 × 10^5	iv	No	D4; D5^p; D8; D11	Blood: ↓ BUN, Cr; kidney: ↓ histology score, TUNEL, ↑ Ki67, ↓ IL-1β, TNFα, IFN-γ, IL-6, TGF-β, MCP-1, ↑ IL-10, bFGF, IGF-1, FIGF, ≈HGF, TGFα, VEGF-A, VEGF-B, VEGF-C, SDF-1; D11: all groups returned to baseline level (BUN, Cr, histology)	Lipophilic carbocyanine dye DIO: 5 min, 30 min; D1, D5: found in the lungs, liver, kidney; D11: not found in the kidney	[14]
Sprague-Dawley	D0: 10 mg/kg ^M4–7	hAd	D1: 5 × 10^5 or; D1: CM 4 ml; D2: CM ml	iv; ip	no	D3	Blood: ↓ BUN, Cr; kidney: ↓ histology score, TUNEL, ↓ Bax, Casp9, Casp3, p-p53, p-ERK, p-JNK, TNFα, COX-2, p-IκB, ≈p-p38; D10: survival (20% versus 0%); CM improved BUN, Cr, histology	BrdU: D3: rarely within the tubular epithelium, also in the lung, not in the liver	[15]
BALB/c; n = 17–60	D0: 18 mg/kg, ip	hUCB; Mouse BM	D1: CM 0.5 ml once or repeated	iv	nr	D4	No effect; Blood: ≈BUN, Cr, histology score, ↓ BW	No	[17]
BALB/cOlaHsd; Immunosuppressed with ATG	D0: 17 mg/kg, ip	hUCB	D1: 5 × 10^5 in 0.2 ml	iv	Yes	D4; D14	MSCs without ATG pretreatment had no effect; MSCs with ATG pretreatment improved survival and renal functional and structural parameters; Blood: ≈BUN, ↓ Cr; kidney: ↓ histology score, ≈casp-3; ↑ HO-1, GPx, ↓ SOD-1, SAA3	Dil: D2: observed mostly in the lungs and liver, rarely in the kidney and intestine	[80]
Athymic nude rats; RNU (lack of T cells); Crl: NIH-Foxn1^rnu	D0: 7 mg/kg, ip	Human kidney-derived cells; CD133^+, CD24^+, CD133^−	D2:10^6 in 0.5 ml PBSs; (and D7: 10^6 in 0.5 ml PBSs)	iv	No	D7	Blood: ↓ BUN, Cr, GFR; kidney: ↓ histology score (no fibrotic lesions); GFR, FITC-sinistrin (D2: increase in FITC in 62% of rats—only these rats were used for the subsequent study); D14: all groups returned to the baseline, regardless of the treatment; second injection did not improve situation; Macrophages in lungs cluster around transplanted cells, ↑ IL10	PKH26 versus GFP: D14: PKH26 cells in kidney close to tubular or interstitial cells and lungs, no evidence of GFP^+ cells; GFP^+ cells were located in the lungs and had disappeared by 24 h	[18]
BALB/c nude	D0: 10 mg/kg,; D1: 10 mg/kg, ip	hAd (third party); HIF-1α modified	D2: 1 × 10^5 in 0.2 ml saline	iv		D5	Blood: ↓ BUN, Cr; kidney: ↓ histology score, ↓ TUNEL, RANTES, ↓ TNFα, ↑ IL-10; PCR: ↑ HO-1; MSCs show protection, but HIF1ɑ-MSCs show greater impact on renal inflammatory factors and HO-1	GFP versus Cy3-labeled CK-18: rare cells overlapped showing rare differentiation of MSC into renal tubular cells CD18^+	[19]
NOD.CB17-Prkdc scid/NcrCrl	D0: 13 mg/kg, sc	hBM reprogram into renal proximal tubular-like cells CL17	D1: 5 × 10^5	iv	No	D4	Blood: ≈ BUN; kidney: ↓ histology score (hyaline casts and necrotic tubuli); Only slight amelioration observed.; CL17 engrafted into proximal tubuli; BM MSCs found mainly at the peritubular level	hMit, cenp-a, PKH26, marker for human mitochondria and centromere protein-A; D4: tubular compartment of the kidney	[20]
NOD-SCID (Charles River)	D0: 13.9 mg/kg, sc	hiPSC-derived renal progenitor cells (RPC); iPSC versus RPC	D1: 5 × 10^5	iv		D4	Blood: ↓ BUN (55%); kidney: ↓ histology score, Ki-67; RPC found in proximal tubuli and rarely in the liver, lung, and spleen, 24 h and D4 after administration; iPSC failed to exert any protective effects not found in kidney or other organs 24 h after administration; 8 wk: teratoma formation and analysis (; 10^6/site; sc)	PKH26, human mitochondria, human nuclear antigen (HNA): 24 h and D4—kidney, liver, lung, heart, and spleen	[21]
BALB/c nude	D0: 14 mg/kg, sc	hUC-derived unrestricted somatic stem cells	D1: 10^5 in 0.5 ml PBS	iv	No	D4	Blood: ↑ BUN, Cr; kidney: ↑ histology score, ~TGF-β1, HGF, IGF-1,; ↓ VEGF-A; IFN-γ, and TNFα—not detectable in some samples; D4: 3 mice died (control group), 3 mice lethargic (treated group); Worsens kidney damage	GFP^+ cells (flow cytometry, IHC): D4: spleen, liver (not detected); lung (0.23% cells were labeled)	[22]
BALB/c nude	D0: 18 mg/kg, iv	hESC; VEGF-modified	D1: 5 × 10^5 in 0.5 ml	iv		D4; M2	Blood: ↓ BUN, Cr; kidney: ↓ TUNEL, histology score, ↑ PCNA, renal capillary density (CD34^+), survival (D14; D9 critical: 60% versus 40% versus 20%); VEGF-MSCs even sign improved versus MSCs; M2: No tumor detected	PKH-26: D4: small number in kidney, large number in the liver, lungs, and spleen	[23]
NOD-SCID; n = 14–17	D0: 11 mg/kg, ip; 6 h: food and water removal	hBM	D1: 5 × 10^6 in 0.37 ml; RPMI	ip		D5^p; (D8D11D15)	Blood: ↓ BUN, Cr, ALT, amylase, phosphorous, ↓ MIP-2, G-CSF, KC, IL-1α, MCP-1, IFN-γ, GM-CSF, IL-6; kidney: ↑ Ki-67, ↓ casp3; D8: BUN returns to the baseline levels; D15: ↑survival (47% versus 10%), 2/7 mice no signal for MSCs engraftment, the same of which the BUN was not affected or not as reduced by the hMSCs	PCR specific for human 1171 bp chromosome 17-specific α-satellite fragment: D5: confirmed in kidney	[24]
NOD-SCID	D0: 12.7 mg/kg, sc	hCB	D1: 5 × 10^6	iv		D4	Blood: ↓ BUN; kidney: ↓ histology score, TUNEL, PCNA, peroxynitrite (oxidative stress), pAkt; EM: peritubular microvascular capillary changes; Survival D9–14; D40 (): no signs of maldifferentiation of MSCs	PKH-26: D4: peritubular areas (83%), in the context of tubular epithelium (5%) and glomeruli (12%), liver, lung, and spleen, rare or absent	[25]

hMSC minimal criteria for defining multipotent MSCs for both laboratory-based scientific investigations and for preclinical studies proposed by the International Society for Cellular Therapy [26]. N: number of animals per group; h: human; D: day; MC: minimal criteria; BM: bone marrow; Ad: adipose tissue; AF: amniotic fluid; UCB: umbilical cord blood; UC: umbilical cord; CB: cord blood; ESC: embryonic stem cells; iPSC: induced pluripotent stem cells; CM: culture media; GFP: green fluorescent protein reporter gene. Unsorted BM MSCs: mixture of hematopoietic cells, mesenchymal stem cells, and lymphoid and myeloid progenitors. sc: subcutaneously; rsc: subcapsular; ia: intra-arterially; iv: intravenously; TUNEL: transferase-mediated dUTP nick end labeling assay; PCNA: proliferating cell nuclear antigen; pFUS: pulsed focused ultrasound; EMT: epithelial-mesenchymal transition; PKH26: lipophilic fluorescent dye; GFR: glomerulat filtration rate; FITC: fluorescein isothiocyanate (FITC)-sinistrin, (a molecule that is filtered by the kidneys, as a measure of the GFR); EM: electron microscopy; IHC: immunohistochemistry; FISH: fluorescent in situ hybridization technique of the human chromosomes; BW: body weight; mALB: microalbumin; β2 mG: macroglobulin; HIF-1α: hypoxia inducible factor-1α; p-peak (of BUN and Cr levels); ATG: polyclonal antithymocyte globuline.