Quantitative analysis of survival of transplanted smooth muscle cells with real-time polymerase chain reaction

Yasuda, Tamotsu; Weisel, Richard D.; Kiani, Chris; Mickle, Donald A.G.; Maganti, Manjula; Li, Ren‐Ke

doi:10.1016/j.jtcvs.2004.06.027

Cited by 61 publications

(57 citation statements)

References 24 publications

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“…We chose biochemical strategies rather than a histological assay, because of the potential for high precision and rapid turnaround [13,38,41,88]. (We recognize the loss of structural information using biochemistry, but this can be obtained in follow up validation studies.)…”

Section: Development Of Rapid Assays For Graft Sizementioning

confidence: 99%

“…These studies showed that a panoply of cells injected into the myocardium both can change the nature of the scar and improve post-infarction cardiac function. Skeletal myoblasts [6][7][8], fetal or neonatal cardiomyocytes [9][10][11], fibroblasts [12], smooth muscle cells [13], hematopoietic stem cells [14,15], mesenchymal stem cells [16], endothelial cells [17], resident cardiac progenitor cells [18,19], and derivatives of human embryonic stem cells [20][21][22][23][24] have all been transplanted into animal hearts. While it is clear that functional benefit is conferred from numerous cell types, the mechanism of action is unclear.…”

Section: Introductionmentioning

confidence: 99%

“…Using smooth muscle cells Yasuda et al reported viable graft cell counts of 15% at 1 week and 9% at 4 weeks after permanent LAD ligation in a rat infarction model [13,42]. Similarly, Hayashi et al reported that 6% of unfractionated bone marrow cells survived at 3 days in infarcted rat [13,42], and Freyman et al reported that ~5% of mesenchymal stem cells survived after transplanting into the infarcted porcine heart [43]. Clearly, there is considerable room for improvement, and solving the problem of cell death is central to successful remuscularization of the infarcted heart.…”

Section: Introductionmentioning

confidence: 99%

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Systems approaches to preventing transplanted cell death in cardiac repair

Robey

Saiget

Reinecke

et al. 2008

Journal of Molecular and Cellular Cardiology

367

294

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Stem cell transplantation may repair the injured heart, but tissue regeneration is limited by death of transplanted cells. Most cell death occurs in the first few days post-transplantation, likely from a combination of ischemia, anoikis and inflammation. Interventions known to enhance transplanted cell survival include heat shock, over-expressing anti-apoptotic proteins, free radical scavengers, anti-inflammatory therapy and co-delivery of extracellular matrix molecules. Combinatorial use of such interventions markedly enhances graft cell survival, but death still remains a significant problem. We review these challenges to cardiac cell transplantation and present an approach to systematically address them.Most anti-death studies use histology to assess engraftment, which is time-and labor-intensive. To increase throughput, we developed two biochemical approaches to follow graft viability in the mouse heart. The first relies on LacZ enyzmatic activity to track genetically modified cells, and the second quantifies human genomic DNA content using repetitive Alu sequences. Both show linear relationships between input cell number and biochemical signal, but require correction for the time lag between cell death and loss of signal. Once optimized, they permit detection of as few as 1 graft cell in 40,000 host cells. Pro-survival effects measured biochemically at three days predict long-term histological engraftment benefits. These methods permitted identification of carbamylated erythropoietin (CEPO) as a pro-survival factor for human embryonic stem cell-derived cardiomyocyte grafts. CEPO's effects were additive to heat shock, implying independent survival pathways. This system should permit combinatorial approaches to enhance graft viability in a fraction of the time required for conventional histology.

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Section: Development Of Rapid Assays For Graft Sizementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Systems approaches to preventing transplanted cell death in cardiac repair

Robey

Saiget

Reinecke

et al. 2008

Journal of Molecular and Cellular Cardiology

367

294

View full text Add to dashboard Cite

show abstract

“…The hydrogels were also shown to exhibit high cell encapsulation efficiencies, supporting cell viability over a one week culture period. These results suggest that the materials appear to be promising to investigate as cell carriers in regenerative medicine approaches, for instance as vehicles to deliver smooth muscle cells for myocardium cell therapy 26 and chondrocyte for cartilage regeneration, 27 although evaluation of specific cell types for cytocompatibility evaluation would be necessary.…”

Section: Hydrogel Degradation and Cytocompatibilitymentioning

confidence: 99%

Protein-Reactive, Thermoresponsive Copolymers with High Flexibility and Biodegradability

Guan

Hong

et al. 2008

Biomacromolecules

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A family of injectable, biodegradable, and thermosensitive copolymers based on Nisopropylacrylamide, acrylic acid, N-acryloxysuccinimide, and a macromer polylactidehydroxyethyl methacrylate were synthesized by free radical polymerization. Copolymers were injectable at or below room temperature and formed robust hydrogels at 37 °C. The effects of monomer ratio, polylactide length, and AAc content on the chemical and physical properties of the hydrogel were investigated. Copolymers exhibited lower critical solution temperatures (LCSTs) from 18 to 26 °C. After complete hydrolysis, hydrogels were soluble in phosphate buffered saline at 37 °C with LCSTs above 40.8 °C. Incorporation of type I collagen at varying mass fractions by covalent reaction with the copolymer backbone slightly increased LCSTs. Water content was 32-80% without collagen and increased to 230% with collagen at 37 °C. Hydrogels were highly flexible and relatively strong at 37 °C, with tensile strengths from 0.3 to 1.1 MPa and elongations at break from 344 to 1841% depending on NIPAAm/HEMAPLA ratio, AAc content, and polylactide length. Increasing the collagen content decreased both elongation at break and tensile strength. Hydrogel weight loss at 37 °C was 85-96% over 21 days and varied with polylactide content. Hydrogel weight loss at 37 °C was 85-96% over 21 days and varied with polylactide content. Degradation products were shown to be noncytotoxic. Cell adhesion on the hydrogels was 30% of that for tissue culture polystyrene but increased to statistically approximate this control surface after collagen incorporation. These newly described thermoresponsive copolymers demonstrated attractive properties to serve as cell or pharmaceutical delivery vehicles for a variety of tissue engineering applications.

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“…12, 14 Briefly, DNA of the whole hearts from groups M, M + R, and M + R + AG (n=4 animals per group) was extracted using the Genomic DNA Purification Kit (Promega), and the concentration of the purified DNA was determined by spectrophotometry. The specific primers were designed according to the GeneBank sequences of the sex-determining region of Y-chromosome (SRY) gene of the rat (sense 5'-CATCGAAGGGTTAAAGTGCCA-3', anti-sense 5'-ATA-GTGTGTAGGTTGTTGTCC-3').…”

Section: Analysis Of Donor Cell Marker and Survivalmentioning

confidence: 99%

Rosuvastatin Treatment Activates JAK-STAT Pathway and Increases Efficacy of Allogeneic Mesenchymal Stem Cell Transplantation in Infarcted Hearts

Yang

Qian

et al. 2011

Circ J

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Quantitative analysis of survival of transplanted smooth muscle cells with real-time polymerase chain reaction

Abstract: One hour after implantation, only 50% of smooth muscle cells remained in the implanted area. Some implanted cells deposited in other tissue. Implanted cell survival progressively decreased during the 4-week study.

Cited by 61 publications

References 24 publications

Systems approaches to preventing transplanted cell death in cardiac repair

Systems approaches to preventing transplanted cell death in cardiac repair

Protein-Reactive, Thermoresponsive Copolymers with High Flexibility and Biodegradability

Rosuvastatin Treatment Activates JAK-STAT Pathway and Increases Efficacy of Allogeneic Mesenchymal Stem Cell Transplantation in Infarcted Hearts

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