Prolonged human/sheep cellular chimerism following transplantation of human hemopoietic stem cellsinto the ewe celomic cavity

Michelini, Monica; Papini, Sandra; Rosellini, Alfredo; Noia, G.; Ligato, Maria Serena; Mancuso, Stefano; Cavazzana, Andrea; Bertacca, Gloria; Cristofano, Claudio Di; Saccardi, Riccardo; Urbani, Serena; Revoltella, Roberto P.

doi:10.1387/ijdb.072362mm

Cited by 6 publications

(4 citation statements)

References 20 publications

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“…Fluorescence in situ hybridization and PCR of the specific sex determining region DNA. To further confirm the successful engraftment of xenotranplanted cells in our model, fluorescence in situ hybridization (FISH) assays for the chromosome Y were performed in female rat hearts that received male pig Cs, as previously described by us (33) and by others (9,45). Briefly, 4-m-thick LV formalin-fixed sections were placed on glass slides pretreated with 3-aminopropyltriethoxysilane and then allowed to dry.…”

Section: Methodsmentioning

confidence: 99%

Ferritin as a reporter gene for in vivo tracking of stem cells by 1.5-T cardiac MRI in a rat model of myocardial infarction

Campan

Lionetti

Aquaro

et al. 2011

American Journal of Physiology-Heart and Circulatory Physiology

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The methods currently utilized to track stem cells by cardiac MRI are affected by important limitations, and new solutions are needed. We tested human ferritin heavy chain (hFTH) as a reporter gene for in vivo tracking of stem cells by cardiac MRI. Swine cardiac stem/progenitor cells were transduced with a lentiviral vector to overexpress hFTH and cultured to obtain cardiospheres (Cs). Myocardial infarction was induced in rats, and, after 45 min, the animals were subjected to intramyocardial injection of ∼200 hFTH-Cs or nontransduced Cs or saline solution in the border zone. By employing clinical standard 1.5-Tesla MRI scanner and a multiecho T2* gradient echo sequence, we localized iron-accumulating tissue only in hearts treated with hFTH-Cs. This signal was detectable at 1 wk after infarction, and its size did not change significantly after 4 wk (6.33 ± 3.05 vs. 4.41 ± 4.38 mm(2)). Cs transduction did not affect their cardioreparative potential, as indicated by the significantly better preserved left ventricular global and regional function and the 36% reduction in infarct size in both groups that received Cs compared with control infarcts. Prussian blue staining confirmed the presence of differentiated, iron-accumulating cells containing mitochondria of porcine origin. Cs-derived cells displayed CD31, α-smooth muscle, and α-sarcomeric actin antigens, indicating that the differentiation into endothelial, smooth muscle and cardiac muscle lineage was not affected by ferritin overexpression. In conclusion, hFTH can be used as a MRI reporter gene to track dividing/differentiating stem cells in the beating heart, while simultaneously monitoring cardiac morpho-functional changes.

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Section: Methodsmentioning

confidence: 99%

Ferritin as a reporter gene for in vivo tracking of stem cells by 1.5-T cardiac MRI in a rat model of myocardial infarction

Campan

Lionetti

Aquaro

et al. 2011

American Journal of Physiology-Heart and Circulatory Physiology

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“…STRs amplified once showed a range of sensitivity varying from 0.1 ng (D8S1179) to 10 ng (D18S51 and D21S11) of human DNA in 1 lg of mouse liver DNA. Nested PCR (see supporting information Table 2) improved sensitivity by a fourth order of magnitude [32].…”

Section: Microsatellite Dna (Short Tandem Repeats)mentioning

confidence: 99%

Human Cord Blood CD133+ Stem Cells Transplanted to Nod-Scid Mice Provide Conditions for Regeneration of Olfactory Neuroepithelium After Permanent Damage Induced by Dichlobenil

et al. 2009

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The herbicide dichlobenil selectively causes necrosis of the dorsomedial part of olfactory neuroepithelium (NE) with permanent damage to the underlying mucosa, whereas the lateral part of the olfactory region and the nasal respiratory mucosa remain undamaged. We investigated here whether human umbilical cord blood CD133 1 stem cells (HSC) injected intravenously to nod-scid mice pretreated with dichlobenil may engraft the olfactory mucosa and contribute to the regeneration of the damaged NE. We tested HLA-DQa1 DNA and three human microsatellites (Combined DNA Index System) as indicators of engrafted cells, finding polymerase chain reaction evidence of chimaerism in various tissues of the host, including the olfactory mucosa and bulb, at 7 and 31 days following HSC transplantation. Histology, immunohistochemistry, and lectin staining revealed the morphological recovery of the dorsomedial region of the NE in dichlobenil-treated mice that received HSC, contrasting with the lack of regeneration in similarly injured areas as these remained damaged in control nontransplanted mice. FISH analysis, to detect human genomic sequences from different chromosomes, confirmed persistent engraftment of the regenerating olfactory area with chimeric cells. Electro-olfactograms in response to odorants, to test the functionality of the olfactory NE, confirmed the functional damage of the dorsomedial area in dichlobenil-treated mice and the functional recovery of the same area in transplanted mice. These findings support the concept that transplanted HSC migrating to the damaged olfactory area provide conditions facilitating the recovery from olfactory receptor cell loss.

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“…The two STRs demonstrated different PCR sensitivity, from 0.1 ng (D8S1179) to 10 ng (D18S51) of human DNA in 1 mg of mouse liver DNA. To improve sensitivity, nested PCR was performed as previously described [28].…”

Section: Polymerase Chain Reaction Analysismentioning

confidence: 99%

Human mesenchymal stromal cell therapy for damaged cochlea repair in nod-scid mice deafened with kanamycin

et al. 2018

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Prolonged human/sheep cellular chimerism following transplantation of human hemopoietic stem cellsinto the ewe celomic cavity

Cited by 6 publications

References 20 publications

Ferritin as a reporter gene for in vivo tracking of stem cells by 1.5-T cardiac MRI in a rat model of myocardial infarction

Ferritin as a reporter gene for in vivo tracking of stem cells by 1.5-T cardiac MRI in a rat model of myocardial infarction

Human Cord Blood CD133+ Stem Cells Transplanted to Nod-Scid Mice Provide Conditions for Regeneration of Olfactory Neuroepithelium After Permanent Damage Induced by Dichlobenil

Human mesenchymal stromal cell therapy for damaged cochlea repair in nod-scid mice deafened with kanamycin

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