Establishment of a complex skin structure via layered co-culture of keratinocytes and fibroblasts derived from induced pluripotent stem cells

Abstract

BackgroundSkin is an organ that plays an important role as a physical barrier and has many other complex functions. Skin mimetics may be useful for studying the pathophysiology of diseases in vitro and for repairing lesions in vivo. Cord blood mononuclear cells (CBMCs) have emerged as a potential cell source for regenerative medicine. Human induced pluripotent stem cells (iPSCs) derived from CBMCs have great potential for allogenic regenerative medicine. Further study is needed on skin differentiation using CBMC-iPSCs.MethodsHuman iPSCs were generated from CBMCs by Sendai virus. CBMC-iPSCs were differentiated to fibroblasts and keratinocytes using embryonic body formation. To generate CBMC-iPSC-derived 3D skin organoid, CBMC-iPSC-derived fibroblasts were added into the insert of a Transwell plate and CBMC-iPSC-derived keratinocytes were seeded onto the fibroblast layer. Transplantation of 3D skin organoid was performed by the tie-over dressing method.ResultsEpidermal and dermal layers were developed using keratinocytes and fibroblasts differentiated from cord blood-derived human iPSCs, respectively. A complex 3D skin organoid was generated by overlaying the epidermal layer onto the dermal layer. A humanized skin model was generated by transplanting this human skin organoid into SCID mice and effectively healed skin lesions.ConclusionsThis study reveals that a human skin organoid generated using CBMC iPSCs is a novel tool for in-vitro and in-vivo dermatologic research.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0958-2) contains supplementary material, which is available to authorized users.

Recapitulating T cell infiltration in 3D psoriatic skin models for patient-specific drug testing

“…However, to apply this approach for other autoimmune diseases and graft-versus-host disease, it will be necessary to consider using donor-matched keratinocytes, fibroblasts, and immune cells. In that case, iPSC-derived skin cells will be an important alternative to address practical difficulties in obtaining multiple tissue types from the same donor 23,50,51 . In addition, considering that other skin components, such as hair follicles 52 , vasculature 24 and adipose tissue 53 also play a role in the regulation of immune reactions in the skin, these components can be included in future constructs using recently developed tissue engineering approaches by our group 21,24 and others 54 .…”

Bioactive Molecules for Skin Repair and Regeneration: Progress and Perspectives

“…RA was a stage-dependent agent for differentiation which directed pluripotent stem cells differentiating into ectodermal cells and upregulated K8/18 and p63 expression [37]. Therefore, RA and BMP4 play distinct but synergistic roles in promoting iPSC or ESC differentiation along the keratinocyte lineage and inhibit neural differentiation [32,38]. Additionally, studies showed that BMP4 and RA promoted the differentiation of pluripotent stem cells into the epidermal through inhibition of the canonical Wnt signaling pathway.…”

Haematopoietic stem cell reprogramming and the hope for a universal blood product

“…By 2008, several groups had adopted the technique, generating iPSCs that were considered epigenetically [44] and developmentally [45] indistinguishable from ESCs of murine [46], nonhuman primate (NHP) [47] and human origin [48,49]. To this day, pluripotent stem cell approaches to regenerative medicine have been proposed to address afflictions from cystic fibrosis [51][52][53] to diabetes [54][55][56] to third-degree burns [57][58][59]. To this day, pluripotent stem cell approaches to regenerative medicine have been proposed to address afflictions from cystic fibrosis [51][52][53] to diabetes [54][55][56] to third-degree burns [57][58][59].…”

“…Concern that cMyc would predispose iPSC-derived cells to malignant transformation was partially quashed when the TF was found to be dispensable to attain pluripotency [50]. To this day, pluripotent stem cell approaches to regenerative medicine have been proposed to address afflictions from cystic fibrosis [51][52][53] to diabetes [54][55][56] to third-degree burns [57][58][59].…”