A modified culture system for epidermal cells for grafting purposes: an in vitro and in vivo study

Dorp, A.G.M. van; Verhoeven, Mary C.H.; Meij, Tineke H. van der Nat–van der; Koerten, Henk K.; Ponec, Maria

doi:10.1046/j.1524-475x.1999.00214.x

Cited by 18 publications

(2 citation statements)

References 31 publications

(31 reference statements)

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“…The human body contains many stem cells, i.e. hematopoietic (HSC) [8], neural [9], epithelial [10, 11] and embryonic stem cells [12]. MSCs do not fulfill all true stem cell criteria.…”

Section: Introductionmentioning

confidence: 99%

Human Bone Marrow-Derived Mesenchymal Stem Cells

et al. 2007

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Mesenchymal stem cells (MSCs) have elicited a great clinical interest, particularly in the areas of regenerative medicine and induction of tolerance in allogeneic transplantation. Previous reports demonstrated the feasibility of transplanting MSCs, which generates new prospects in cellular therapy. Recently, injection of MSCs induced remission of steroid-resistant acute graft-versus-host disease (GVHD). This review summarizes the knowledge and possible future clinical uses of MSCs.

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“…The human body contains many stem cells, i.e. hematopoietic (HSC) [8], neural [9], epithelial [10, 11] and embryonic stem cells [12]. MSCs do not fulfill all true stem cell criteria.…”

Section: Introductionmentioning

confidence: 99%

Human Bone Marrow-Derived Mesenchymal Stem Cells

et al. 2007

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“…5,24 The carrier was porous scaffold made from biodegradable poly(ethyleneglycol terephthalate)-poly(butylene terephthalate) copolymers (PEGT/PBT). 5,25 Previously, it was demonstrated to be an appropriate growth substrate for skin cells [26][27][28][29][30] and to support connective tissue repair in rat and porcine wound models. 31,32 The objectives in this study were to evaluate differences between human fibroblasts isolated from adipose tissue and papillary dermis and their role in the repair of full-thickness wounds in athymic mice.…”

Section: Introductionmentioning

confidence: 99%

Stimulation of Skin Repair Is Dependent on Fibroblast Source and Presence of Extracellular Matrix

Wang

Pieper²,

Schotel³

et al. 2004

Tissue Engineering

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In this study in vitro and in vivo functions were compared between cultured dermal equivalents produced with human fibroblasts isolated either from papillary dermis or adipose tissue of the same donors. Papillary dermal fibroblasts had a normal spindle cell shape; in contrast, adipose tissue fibroblasts had a stellate cell shape, actin stress fibers containing alpha-smooth muscle actin, multiple narrow extensions at their edges, and longer focal adhesion plaques. After dynamic culture for 14 days in PEGT/PBT carrier scaffolds, cell numbers between the two cell sources were comparable, but tissue morphology was different between the cultured groups. In addition, papillary fibroblasts had deposited significantly more glycosaminoglycans (214 +/- 15 versus 159 +/- 21 microg, p < 0.001) and a lower amount of collagen (49 +/- 14 versus 111 +/- 25 microg of hydroxyproline, p < 0.001) than had adipose fibroblasts. Moreover, the latter constructs were significantly more contracted than the papillary fibroblast-cultured constructs (78 +/- 6 versus 96 +/- 3%, p < 0.001). In comparison with the influence of cultured dermal equivalents on wound healing, the transplantation of five groups (control acellular carrier, papillary fibroblast-seeded construct, adipose fibroblast-seeded construct, papillary fibroblast-cultured construct, and adipose fibroblast-cultured construct) to full-thickness wounds on the backs of athymic mice showed clear differences in angiogenesis and tissue ingrowth after 10 days, and in reepithelialization after 21 days. After 10 days, the level of vascular ingrowth in the carrier (von Willebrand staining) for the five groups was as follows: adipose fibroblast-cultured > papillary fibroblast-cultured = adipose fibroblast-seeded > papillary fibroblast-seeded > acellular carrier. After 21 days, only the acellular carriers were not vascularized and the papillary fibroblast-seeded constructs were not completely vascularized. Complete wound reepithelialization (92 +/- 12%) was observed only in the group treated with adipose cultured constructs. Wound contraction was not observed. Staining for HLA-ABC and alpha-smooth muscle actin showed that human fibroblasts had survived and that adipose fibroblasts continued to express the actin isoform. These results showed not only stimulation of skin repair when fibroblasts were present in the carrier, but also significant positive effects of the deposited extracellular matrix (ECM) in the carrier. In addition, the adipose fibroblast-seeded construct, and especially the adipose fibroblast-cultured construct, significantly stimulated angiogenesis and reepithelialization when compared with their corresponding papillary fibroblast constructs. Apparently, tissue source or fibroblast phenotype and the presence of ECM play a crucial role in the stimulation of (impaired) healing and engineering of dermal equivalents.

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Synthetic scaffold morphology controls human dermal connective tissue formation

Wang

Pieper²,

Péters³

et al. 2005

J Biomedical Materials Res

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Engineering tissues in bioreactors is often hampered by disproportionate tissue formation at the surface of scaffolds. This hinders nutrient flow and retards cell proliferation and tissue formation inside the scaffold. The objective of this study was to optimize scaffold morphology to prevent this from happening and to determine the optimal scaffold geometric values for connective tissue engineering. After comparing lyophilized crosslinked collagen, compression molded/salt leached PEGT/PBT copolymer and collagen-PEGT/PBT hybrid scaffolds, the PEGT/PBT scaffold was selected for optimization. Geometric parameters were determined using SEM, microcomputed tomography, and flow permeability measurements. Fibroblast were seeded and cultured under dynamic flow conditions for 2 weeks. Cell numbers were determined using CyQuant DNA assay, and tissue distribution was visualized in H&E- and Sirius Red-stained sections. Scaffolds 0.5 and 1.5 mm thick showed bridged connected tissue from top-to-bottom, whereas 4-mm-thick scaffolds only revealed tissue ingrowth until a maximum depth of 0.6-0.8 mm. Rapid prototyped scaffold were used to assess the maximal void space (pore size) that still could be filled with tissue. Tissue bridging between fibers was only found at fiber distances < or =401 +/- 60 microm, whereas filling of void spaces in 3D-deposited scaffolds only occurred at distances < or =273 +/- 55 microm. PEGT/PBT scaffolds having similar optimal porosities, but different average interconnected pore sizes of 142 +/- 50, 160 +/- 56 to 191 +/- 69 microm showed comparable seeding efficiencies at day 1, but after 2 weeks the total cell numbers were significantly higher in the scaffolds with intermediate and high interconnectivity. However, only scaffolds with an intermediate interconnectivity revealed homogenous tissue formation throughout the scaffold with complete filling of all pores. In conclusion, significant amount of connective tissue was formed within 14 days using a dynamic culture process that filled all void spaces of a PEGT/PBT scaffolds with the following geometric parameters: thickness 1.5-1.6 mm, pore size range 90-360 microm, and average interconnecting pore size of 160 +/- 56 microm.

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A modified culture system for epidermal cells for grafting purposes: an in vitro and in vivo study

Cited by 18 publications

References 31 publications

Human Bone Marrow-Derived Mesenchymal Stem Cells

Human Bone Marrow-Derived Mesenchymal Stem Cells

Stimulation of Skin Repair Is Dependent on Fibroblast Source and Presence of Extracellular Matrix

Synthetic scaffold morphology controls human dermal connective tissue formation

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