Novel Embryoid Body–Based Method to Derive Mesenchymal Stem Cells from Human Embryonic Stem Cells

Abstract

Application of human embryonic stem cells (hESCs) to stem-cell therapy is not feasible because of the risk of tumorigenicity and rejection. In contrast, human mesenchymal stem cells (hMSCs) are free from the risk of tumorigenicity and also have immune privilege. However, hMSCs obtained from adults have infinite variety in terms of the biological characteristics and functionality. We report here a new derivation method of hMSCs from hESCs. The derivation of hMSCs from three different hESC lines (SNUhES3, CHA3-hESC, and H9) was performed by embryoid bodies formation and subsequent culture with stage-different media without using inductive xenogenic feeder and mechanical selection procedure. The derived cells were morphologically similar to the unique fingerprint-like pattern of hMSCs and grew stably for at least 35 passages in vitro. These cells had hMSCs-like immunophenotypes: negative for CD34 and CD45; positive for CD29, CD44, CD73, CD90, and CD105. They could be differentiated into multiple lineages including osteocytes, chondrocytes, adipocytes, and myocytes. They maintained normal karyotype during the long-term cultivation and did not show tumorigenicity when transplanted into the immunodeficient mice. In conclusion, the new embryoid body-based derivation method of hMSCs from hESCs is simple, safe, and reproducible in three different hESC lines. We expect that this method will provide a more effective and powerful tool to derive hMSCs from various hESC lines.

Mapping the first stages of mesoderm commitment during differentiation of human embryonic stem cells

“…The generation from hESCs of several progenitor populations with limited mesodermal potential has been previously described using combinations of markers of hematoendothelial (CD34 and PDGFR-α), cardiovascular (KDR + CD117 − cells), and mesenchymal (CD73) differentiation (3,4,6,21). We now demonstrate that day 3.5 CD326 − CD56 + cells represent the earliest multipotent mesodermal progenitors reported to date, being generated before the appearance of the previously described populations and capable of giving rise to all of the above progenitor cell phenotypes and related mesodermal lineages.…”

Directed Differentiation of Human Induced Pluripotent Stem Cells Toward Bone and Cartilage: In Vitro Versus In Vivo Assays

“…There are reports of attempts to generate osteo-and chondroprogenitors (also known as bone marrow-derived mesenchymal stem cells [MSCs]) from embryonic stem cells (ESCs) [1][2][3][4][5][6][7][8][9][10] and iPSCs [11][12][13][14]. However, there are three broad limitations in performing these types of studies.…”

Human Embryonic Stem Cells Derived Mesenchymal Stem/Stromal Cells and their Use in Regenerative Medicine

“…Recent studies reported that MSCs can be derived from hESCs [9][10][11] that these differentiated cells expressed MSCs surface markers [2,9]. Also, our preliminary data showed hESCs derived MSCs (hESCs-MSCs) with fibroblast-like morphology ( Figure 1) expressed CD105 and FIBRONECTIN as MSC specific markers (unpublished data; Figure 1).…”