Increased Yield of Endothelial Cells from Peripheral Blood for Cell Therapies and Tissue Engineering

Jamiolkowski, Ryan M.; Kang, Sa Do; Rodriguez, AnnMarie K; Haseltine, Justin M.; Galinat, Lauren J.; Jantzen, Alexandra E.; Carlon, Timothy; Darrabie, Marcus D.; Arciniegas, Antonio J.; Mantilla, José G.; Haley, Nancy; Noviani, Maria; Allen, Jason D.; Stäbler, Thomas; Frederiksen, James W.; Álzate, Óscar; Keil, Lukas G.; Liu, Siyao; Lin, Fu-Hsiung; Truskey, George A.; Achneck, Hardean E.

doi:10.2217/rme.15.2

Cited by 8 publications

(6 citation statements)

References 39 publications

(46 reference statements)

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“…Autologous circulating endothelial progenitor cells were isolated and cultured from peripheral mononuclear cells of the oral mucosal samples of each experimental rat using a diluted whole blood incubation technique, as previously described 23 , 39 . Peripheral blood from the same rat was obtained and anticoagulated with 20-USP heparin units/mL blood (Sigma-Aldrich).…”

Section: Methodsmentioning

confidence: 99%

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RETRACTED ARTICLE: Accelerated oral wound healing using a pre-vascularized mucosal cell sheet

Lee

Kim

Shin

et al. 2017

Sci Rep

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Cell sheets with pre-vascularization have recently been developed but remain relatively untested in oral wound healing. Therefore, we examined the potential utility of our newly developed pre-vascularized mucosal cell sheets in oral wound healing. Mucosal keratinocytes, fibroblasts, and endothelial progenitor cells were primarily cultured for in vitro cell expansion from mucosa and blood of Sprague-Dawley rats. Mucosal cell sheets were generated using cultured keratinocytes and plasma fibrin (K sheet) or keratinocytes and a mixture of fibrin, fibroblasts, and endothelial cells (PV sheet). Autologous sheets were transplanted on deep wounds in the buccal region of rats. The gross and histological characteristics of wound healing were compared among control wound, K sheet, and PV sheet groups. We successfully cultured and expanded keratinocytes, fibroblasts, and endothelial progenitor cells in vitro for generating mucosal cell sheets with or without pre-vascularization. In the in vivo oral wound model, compared with the control wound, the PV sheet group exhibited rapid wound closure more prominently than the K sheet group. The histological healing in the PV sheet group was similar to that in rat normal buccal mucosa without fibrosis. The pre-vascularized mucosal cell sheet exhibited in vivo efficacy in oral wound healing by promoting accelerated healing.

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Section: Methodsmentioning

confidence: 99%

“…1 ). Endothelial progenitor cells can be isolated from peripheral blood cells 22 , 23 . The pre-vascularized layer beneath the keratinocyte layer contained pre-formed microvessels generated by isolating and expanding endothelial cells from peripheral blood samples.…”

Section: Introductionmentioning

confidence: 99%

RETRACTED ARTICLE: Accelerated oral wound healing using a pre-vascularized mucosal cell sheet

Lee

Kim

Shin

et al. 2017

Sci Rep

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“…In addition, fibroblasts produce collagen fibres and replace fibrin matrix, contributing to a coherent three-dimensional cell-sheet structure 22 , 23 . Further, the cell sheets included pre-formed microvessels derived from the EPC isolated from adult peripheral blood monocytes as reported previously 17 , 24 . The pre-formed microvessels might have contributed to the early survival of the tissue-engineered cell sheets in the wound bed by ensuring sufficient blood supply, resulting in accelerated wound healing.…”

Section: Discussionmentioning

confidence: 99%

Use of a pre-vascularised oral mucosal cell sheet for promoting cutaneous burn wound healing

2018

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Pre-vascularised cell sheets have been used to promote early angiogenesis and graft survival. However, the use of pre-vascularised mucosal cell sheets for burn wounds has been rarely evaluated. Therefore, we examined the applicability of an oral pre-vascularised mucosal cell sheet that we had previously developed for the treatment of cutaneous burn wounds.Methods: Mucosal keratinocytes, fibroblasts, and endothelial progenitor cells were isolated from the oral mucosa and peripheral blood and were expanded in vitro. Mucosal cell sheets were generated by seeding cultured keratinocytes onto a mixture of fibroblasts, endothelial cells, and fibrin. Third-degree burn wounds were created on the backs of rats and were covered with the cell sheets, skin grafts, or silastic sheets as a control. Gross and microscopic findings and gene expression profiles of wounds were compared among the groups.Results: CD31-positive microvessels were observed in the fibrin-matrix layer of the cell sheet. In the cutaneous burn wound model, the cell sheets promoted wound healing, with accelerated wound closure and less scarring than with silastic sheets and skin grafts. The cell sheets had more microvessels and proliferating cells and less neutrophil infiltration and fibrotic features than the controls or skin grafts. The cell sheet induced higher mRNA expression of KRT14, VEGFA, IL10, and AQP3 and lower mRNA expression of TGFB1, IL6, ICAM1, ACTA2, and FN1 than did the controls or skin grafts.Conclusions: The pre-vascularised mucosal cell sheet promotes cutaneous burn wound healing.

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“…6 We further showed that the oral mucosal cell sheets might have the necessary tissue plasticity for promoting rapid and scarless cutaneous wound healing, with in vivo effects equivalent to skin-derived cell sheets in a rat skin wound model. 9,10 The in vivo examinations of the prevascularized oral mucosal cell sheet might guide the clinicians to potentially use the cell sheet engineering from the oral mucosa in the patients with cutaneous burn wounds. 8 The cell sheets include the preformed microvessels from the endothelial progenitor cells isolated from adult peripheral blood monocyte cells as reported previously.…”

Section: Introductionmentioning

confidence: 99%

“…8 The cell sheets include the preformed microvessels from the endothelial progenitor cells isolated from adult peripheral blood monocyte cells as reported previously. 9,10 The in vivo examinations of the prevascularized oral mucosal cell sheet might guide the clinicians to potentially use the cell sheet engineering from the oral mucosa in the patients with cutaneous burn wounds. Therefore, we examined the potential use of our newly developed prevascularized oral mucosal cell sheet in skin excisional wounds.…”

mentioning

confidence: 99%

Promotion of skin wound healing using prevascularized oral mucosal cell sheet

2018

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Background This study examined the potential use of our newly developed prevascularized oral mucosal cell sheet for the treatment of skin wounds. Methods Mucosal cell sheets containing cultured keratinocytes and plasma fibrin without (K sheet) or with a mixture of fibroblasts and endothelial progenitor cells (PV sheet) were transplanted into full‐thickness skin excisional wounds of nude mice. Results This technique was successful for in vitro culture; expanding keratinocytes, fibroblasts, and endothelial progenitor cells; and generating prevascularized mucosal cell sheets. Cell sheets promoted in vivo wound healing with rapid wound closure and less scarring compared to controls. This result was more apparent in the PV than the K sheet (P < .05). Wounds covered with cell sheets showed less expression of TGFB1, ACTA2, and FN1 mRNAs than the controls (P < .05). Conclusion The prevascularized mucosal cell sheet showed in vivo efficacy and tissue plasticity in cutaneous wounds by promoting accelerated healing.

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Increased Yield of Endothelial Cells from Peripheral Blood for Cell Therapies and Tissue Engineering

Cited by 8 publications

References 39 publications

RETRACTED ARTICLE: Accelerated oral wound healing using a pre-vascularized mucosal cell sheet

RETRACTED ARTICLE: Accelerated oral wound healing using a pre-vascularized mucosal cell sheet

Use of a pre-vascularised oral mucosal cell sheet for promoting cutaneous burn wound healing

Promotion of skin wound healing using prevascularized oral mucosal cell sheet

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