Stable cell adhesion affects mesenchymal stem cell sheet fabrication: Effects of fetal bovine serum and human platelet lysate

Abstract

Cell sheet technology exploits temperature responsive cell culture dishes (TRCDs) as versatile cell harvesting methods to yield contiguous cell monolayers robustly held together by cell-cell junctions, receptors, and endogenous extracellular matrix. More than 15 years of clinical data using autologous-sourced cell sheets demonstrate enhanced therapeutic properties through increased cell retention at target tissue sites. Recently, several preclinical studies have also been reported using mesenchymal stem cell (MSC) sheets in wound healing, cardiac ischemia therapies, and pancreatic regeneration. However, optimized MSC sheet fabrication conditions have not yet been reported. In this study, we identified specific conditions for reliable human MSC sheet fabrication by comparing cell growth media supplements (fetal bovine serum [FBS] and human platelet lysate [hPL]). Human umbilical cord-derived MSCs cultured in FBS and hPL exhibit different actin cytoskeletal structures related to their cell morphologies and adhesion. MSCs cultured in FBS media showed stable cell adhesion on TRCDs with flattened cell shapes and aligned actin cytoskeletal structure. This stable cell adhesion enables production of consistent MSC cell sheets, with controlled cell sheet detachment. Conversely, cell sheet fabrication in hPL media exhibits poor reproducibility being more sensitive to temperature-and culture timeinduced release due to weak cell adhesion. These findings suggest that stable MSC adhesion to TRCDs is important to reliable MSC sheet fabrication methods and that MSC growth media supplementation directly affects cell adhesion during culture. K E Y W O R D S cell adhesion, cell morphology, cell therapy, cytokine secretion, cytoskeleton structure, fetal bovine serum, human platelet lysate

Human Mesenchymal Stem Cell Sheets Improve Uterine Incision Repair in a Rodent Hysterotomy Model

“…18,47,48 Significant HGF and VEGF expression in these human umbilical cord stem cell sheets is already known. 19,51 Previous studies have shown that transformation of transplanted stem cells into myometrial tissue is unlikely to be a significant contributor to repair, at least not in the acute phase. 18 Further, transplantation of allogenic myometrial cells did not improve hysterotomy wound healing in an animal uterine repair model.…”

Measurement of the Adipose Stem Cells Cell Sheets Transmittance

“…It has been shown that transformation of a MSC cell sheet from 2D monolayer to 3D multilayer upon detachment from cell culture dish resulted in morphological rearrangement of actin cytoskeleton from unidirectional to multi-directional alignment, which makes a 3D multilayer cell sheet opaquer [52]. Furthermore, MSC cell sheets fabricated with supplementation of fetal bovine serum (FBS) and human platelet lysate (hPL) exhibited unidirectionally and multi-directionally aligned actin cytoskeletal structures, respectively, where cell sheets with multi-directional actin structure had relatively opaque appearance [53]. It is reasonable to associate the opaqueness of these cell sheets to its multi-directional, random orientation of actin cytoskeleton, as it augments the degree of scattering of irradiated light.…”

Thermoresponsive Cationic Block Copolymer Brushes for Temperature‐Modulated Stem Cell Separation

“…The prepared thermoresponsive block copolymer brush was investigated as a cell separation tool, using umbilical cord‐derived mesenchymal stem cells (UCMSC) as target cells because the UCMSC are used as effective therapeutic cells. [ 37,38 ] In the process of the collection of UCMSC from the umbilical cord, fibroblasts and macrophages tend to mix with the UCMSC. Thus, normal human dermal fibroblasts (NHDFs) and RAW264.7 cells were used as models of the contaminant cells, fibroblasts and macrophages, respectively.…”