Scientific Publications by FDA Staff
Stem Cells Dev 2005 Jun;14(3):270-84
Development of a focused microarray to assess human embryonic stem cell differentiation.
Yang AX, Mejido J, Luo Y, Zeng X, Schwartz C, Wu T, Thies RS, Bhattacharya B, Han J, Freed B, Rao M, Puri RK
Puri RK, US FDA, Lab Mol & Tumor Biol, Div Cellular & Gene Therapies, Ctr Biol Evaluat & Res, NIH Bldg 29B,Room 2NN20,29 Lincoln Dr, Bethesda, MD 20892 USA US FDA, Lab Mol & Tumor Biol, Div Cellular & Gene Therapies, Ctr Biol Evaluat & Res, Bethesda, MD 20892 USA NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA Natl Inst Drug Abuse, Cellular Neurobiol Branch, Baltimore, MD 21224 USA Natl Inst Dent & Craniofacial Res, Biostat Core Unit, NIH, Bethesda, MD 20892 USA Geron Corp, Menlo Pk, CA 94025 USA Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21218 USA
Human embryonic stem cells (hESC) must be differentiated before clinical use. In addition, the extent of contamination of undifferentiated cells and the efficiency of differentiation must also be assessed prior to clinical application. In this manuscript, we describe the development of a focused microarray that may be used to discriminate between hESC and their differentiated progeny. This array contains 755 genes including embryonic stem cell markers as well as markers of differentiation into neural, mesodermal, and endodermal phenotypes. In addition, we have included candidate genes belonging to families of cytokines, chemokines, receptors, signaling pathways, and homeodomain proteins that are likely to be important in the process of differentiation. Testing and validation of the focused array was performed using RNA from hESC, human embryoid body (hEB) outgrowths, and a human embryonal carcinoma (hEC) cell line. We have compared gene expression with negative background, GAPDH, beta-actin positive controls, and human universal RNA (hURNA), showing that such an array can rapidly distinguish between undifferentiated and differentiated hESC-derived cell populations. We expect that the described array will be extremely useful in evaluating the extent of differentiation and the state of the hESC-derived population utilized for therapeutic purposes.
|Category: Journal Article|
|PubMed ID: #15969622|
|Includes FDA Authors from Scientific Area(s): Biologics|
|Entry Created: 2011-10-04||Entry Last Modified: 2012-08-29|