Differentiation Ability of Amniotic Fluid-Derived Stem Cells Cultured on Extracellular Matrix-immobilized Surface

Abstract

Stem cells from amniotic fluid were cultured on dishes coated or grafted with extracellular matrix (ECM) or Matrigel where gelatin, collagen, fibronectin, laminin, and vitronectin were selected as ECM components (nanosegments). The effects of interactions between amniotic fluid stem cells and nanosegments were investigated on the expression of surface markers of mesenchymal stem cells and on the differentiation abilities of osteoblasts and neural cells. The ECM-coated dishes produced water contact angles from 35 to 65 degrees, whereas ECM-grafted dishes produced water contact angles from 50 to 70 degrees, which was an adequate water contact angle range for our cell culture conditions. Culture on ECM-immobilized dishes enhances amniotic fluid stem cell differentiation into osteoblasts more than culture on polystyrene dishes grafted with amino groups (PS-NH2 dishes). This finding indicates that specific interactions between amniotic fluid cells and the ECM grafted onto the culture dishes promote the differentiation of cells into osteoblasts. Matrigel-immobilized dishes promoted a more extensive differentiation of amniotic fluid stem cells into neural cells than did ECMimmobilized dishes, but they did not promote differentiation into osteoblasts. Immobilization of the optimal nanosegments (ECM or Matrigel) onto culture dishes enhances amniotic fluid stem cell differentiation into osteoblasts and neural cells; the choice of nanosegments depends on the desired differentiated cell type.

Transient characteristics of universal cells on human‐induced pluripotent stem cells and their differentiated cells derived from foetal stem cells with mixed donor sources

“…The AF was centrifuged at 255 × g for 300 seconds, and the supernatant was discarded (Figure 1). The AF cells (cell pellets) were suspended in medium composed of MCDB 201/DMEM (60%/40%) supplemented with 10 ng/mL FGF‐2 and 20% foetal bovine serum (FBS) and cultivated on tissue culture polystyrene (TCPS) plates in a CO 2 incubator at 37°C to obtain human amniotic fluid stem cells (hAFSCs) 28 . After reaching approximately 78%‐82% confluence, the cells (hAFSCs) were detached with a 0.25% trypsin‐EDTA solution, centrifuged and inoculated into TCPS dishes according to a conventional passage procedure (Figure 1).…”

“…After reaching approximately 78%‐82% confluence, the cells (hAFSCs) were detached with a 0.25% trypsin‐EDTA solution, centrifuged and inoculated into TCPS dishes according to a conventional passage procedure (Figure 1). 28 hAFSCs at passage 3‐4 were used for the following reprogramming processes.…”

“…The AF cells (cell pellets) were suspended in medium composed of MCDB 201/DMEM (60%/40%) supplemented with 10 ng/mL FGF-2 and 20% foetal bovine serum (FBS) and cultivated on tissue culture polystyrene (TCPS) plates in a CO 2 incubator at 37°C to obtain human amniotic fluid stem cells (hAFSCs). 28 After reaching approximately 78%-82% confluence, the cells (hAFSCs) were F I G U R E 1 Outline of the process of reprogramming hAFSCs into transient universal (hypoimmunogenic) hiPSCs prepared from multiple AF donors (A) and conventional hiPSCs prepared from a single AF donor (B). The immunotolerance of transient universal (hypoimmunogenic) hiPSCs and their differentiated cardiomyocytes or embryoid bodies (EBs) is evaluated after treatment with mononuclear cells donated by different AF donors detached with a 0.25% trypsin-EDTA solution, centrifuged and inoculated into TCPS dishes according to a conventional passage procedure ( Figure 1).…”

“…The immunotolerance of transient universal (hypoimmunogenic) hiPSCs and their differentiated cardiomyocytes or embryoid bodies (EBs) is evaluated after treatment with mononuclear cells donated by different AF donors detached with a 0.25% trypsin-EDTA solution, centrifuged and inoculated into TCPS dishes according to a conventional passage procedure ( Figure 1). 28 hAFSCs at passage 3-4 were used for the following reprogramming processes.…”

Drug-resistant colon cancer cells produce high carcinoembryonic antigen and might not be cancer-initiating cells

“…Conventional staining protocol35–39 was used. The expression of CD133, forward scattering intensity, and side scattering intensity of the LoVo cells were analyzed by flow cytometry (Coulter EPICS™ XL; Beckman-Coulter, Brea, CA, USA).…”