High‐efficient generation of VCAM‐1⁺ mesenchymal stem cells with multidimensional superiorities in signatures and efficacy on aplastic anaemia mice

Abstract

Objective Longitudinal studies have indicated VCAM‐1+ mesenchymal stem/stromal cells (MSCs) as promising resources in regenerative medicine, yet the abundance in gene expression is far from adequate in the advantaged and “discarded” hUC‐MSCs. Thus, high‐efficient preparation and systematic dissection of the signatures and biofunctions of the subpopulation is the prerequisite for large‐scale clinical applications. Materials and methods We primarily took advantage of a cytokine‐based programming strategy for large‐scale VCAM‐1+ hUC‐MSC generation (III‐MSCs). Thereafter, we conducted multifaceted analyses including cytomorphology, immunophenotype, cell vitality, multilineage differentiation, whole‐genome analysis, tube formation and Matrigel plug assay, lymphocyte activation and differentiation, and systemic transplantation for aplastic anaemia (AA) treatment. Results III‐MSCs with high‐proportioned VCAM‐1 expression were obtained by combining IL‐1β, IL‐4 with IFN‐γ, which exhibited comparable immunophenotype with untreated hUC‐MSCs (NT‐MSCs) but revealed multidimensional superiorities both at the cellular and molecular levels. Simultaneously, systemic infusion of III‐MSCs could significantly ameliorate clinicopathological features and finally help facilitate haematopoietic reconstruction and immunoregulation in AA mice. Conclusions We have established a high‐efficient procedure for large‐scale generation of III‐MSCs with preferable signatures and efficacy upon aplastic anaemia in mice. Our findings suggested that III‐MSCs were advantageous sources with multifaceted characteristics for regenerative medicine.

Two-step generation of mesenchymal stem/stromal cells from human pluripotent stem cells with reinforced efficacy upon osteoarthritis rabbits by HA hydrogel

“…The total RNAs for transcriptome analysis were extracted from hESC-MSCs and hiPSC-MSCs and prepared as we recently reported with several modifications [ 5 , 6 , 21 ]. Briefly, 5 × 10 6 hESC-MSCs or hiPSC-MSCs were used for RNA-SEQ analysis by Novogene (Novogene, Tianjin, China) and the bioinformatic analyses were conducted based on the raw data.…”

“…Mesenchymal stem/stromal cells (MSCs), acknowledged as multipotent mesenchymal progenitor/precursor cells, medicinal signaling cells or skeletal stem cells, are heterogeneous populations with hematopoietic supporting and immunoregulatory attributes as well as multilineage differentiation potential [ 1 – 3 ]. We and other investigators have verified the dominant role of MSCs in multiple physiological and pathological microenvironments, together with clinical and preclinical applications in regenerative medicine such as osteoarthritis [ 4 ], aplastic anemia [ 5 , 6 ], Crohn’s disease [ 7 ], immune thrombocytopenia [ 8 ], fulminant hepatic failure [ 9 ], acute myocardial infarction (AMI) [ 10 ], graft-versus-host disease (GVHD) [ 11 ] and even the current corona virus disease 2019 (COVID-19) [ 12 ] through direct- and trans-differentiation, autocrine and paracrine, targeting immune-regulating cells and repairing damaged tissues. Since the 1960s, MSCs have been successfully isolated and identified from extensive sources including adult tissues (bone marrow, dental pulp, adipose and liver tissue) and perinatal tissues (placenta, umbilical cord, amniotic fluid and membrane) [ 13 – 16 ].…”

Interferon-γ enhances the therapeutic effect of mesenchymal stem cells on experimental renal fibrosis

“…High mobility group box-1 protein (HMGB1) and interleukin-18 (IL-18) are members of DAMPs— HMGB1 was reported to promote the migration of MSCs 19 , 20 , whereas IL-18 contributed to the secretion of interferon-γ (IFN-γ) from natural killer cells 21 . Furthermore, IFN-γ released from immune cells at sites of damaged tissues stimulated MSCs to secrete anti-inflammatory mediators including prostaglandin E2 (PGE2) 22 – 26 . PGE2 has been reported to induce the polarization of immunosuppressive M2 macrophages that produce anti-inflammatory cytokines and inhibit the persistence of inflammation 27 , 28 .…”

Multifaceted modifications for a cell size‐based circulating tumor cell scope technique hold the prospect for large‐scale application in general populations

“…The immunofluorescence (IF) staining was performed as described before with several modifications (Wei et al, 2019, 2020; Zhang, Wang, et al, 2018). Briefly, the fixed cells on the filter membrane were incubated in 4% formaldehyde for 10 min and blocked with the blocking buffer for 1 h (ScreenCell).…”