Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies

Sergey Malchenko; Jianping Xie; Maria de Fatima Bonaldo; Elio F. Vanin; Bula J. Bhattacharyya; Abdelhak Belmadani; Guifa Xi; Vasily Galat; William Goossens; Richard E.B. Seftor; Tadanori Tomita; John Crispino; Richard J. Miller; Martha C. Bohn; Mary J.C. Hendrix; Marcelo B. Soares

doi:10.1016/j.gene.2013.07.101

Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies

Sergey Malchenko, Jianping Xie, Maria de Fatima Bonaldo, Elio F. Vanin, Bula J. Bhattacharyya, Abdelhak Belmadani, Guifa Xi, Vasily Galat, William Goossens, Richard E.B. Seftor, Tadanori Tomita, John Crispino, Richard J. Miller, Martha C. Bohn, Mary J.C. Hendrix, Marcelo B. Soares^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

In vitro neural differentiation of human embryonic stem cells (hESCs) is an advantageous system for studying early neural development. The process of early neural differentiation in hESCs begins by initiation of primitive neuroectoderm, which is manifested by rosette formation, with consecutive differentiation into neural progenitors and early glial-like cells. In this study, we examined the involvement of early neural markers - OTX2, PAX6, Sox1, Nestin, NR2F1, NR2F2, and IRX2 - in the onset of rosette formation, during spontaneous neural differentiation of hESC and human induced pluripotent stem cell (hiPSC) colonies. This is in contrast to the conventional way of studying rosette formation, which involves induction of neuronal differentiation and the utilization of embryoid bodies. Here we show that OTX2 is highly expressed at the onset of rosette formation, when rosettes comprise no more than 3-5 cells, and that its expression precedes that of established markers of early neuronal differentiation. Importantly, the rise of OTX2 expression in these cells coincides with the down-regulation of the pluripotency marker OCT4. Lastly, we show that cells derived from rosettes that emerge during spontaneous differentiation of hESCs or hiPSCs are capable of differentiating into dopaminergic neurons in vitro, and into mature-appearing pyramidal and serotonergic neurons weeks after being injected into the motor cortex of NOD-SCID mice.

Original language	English (US)
Pages (from-to)	400-407
Number of pages	8
Journal	Gene
Volume	534
Issue number	2
DOIs	https://doi.org/10.1016/j.gene.2013.07.101
State	Published - Jan 25 2014

Funding

This work was supported in part by the Ann & Robert H Lurie Children's Hospital of Chicago Research Center “Excellence in Academic Medicine” Grant from the Illinois Department of Public Aid (Soares, PI) , the Dr. Ralph and Marian C. Falk Medical Research Trust , the Children Research Fund, the Gus Foundation , the Maeve McNicholas Memorial Foundation , the Everett/O'Connor Charitable Trust , NIH/NCI CA121205 (Hendrix, PI), the Medical Research Institute Council (Bohn), the Northwestern University Flow Cytometry Facility a Cancer Center Support Grant ( NCI CA060553 ), and NIH ARRA Grant RC1HL100168 (Crispino, PI).

Keywords

Induced pluripotent stem cells
Mature pyramidal neurons
Neural rosettes
Radial glia

ASJC Scopus subject areas

Genetics

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10.1016/j.gene.2013.07.101

Cite this

Malchenko, S., Xie, J., de Fatima Bonaldo, M., Vanin, E. F., Bhattacharyya, B. J., Belmadani, A., Xi, G., Galat, V., Goossens, W., Seftor, R. E. B., Tomita, T., Crispino, J., Miller, R. J., Bohn, M. C., Hendrix, M. J. C., & Soares, M. B. (2014). Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies. Gene, 534(2), 400-407. https://doi.org/10.1016/j.gene.2013.07.101

@article{cbc4e03d2a67449ea6ce32a920e62e7e,

title = "Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies",

abstract = "In vitro neural differentiation of human embryonic stem cells (hESCs) is an advantageous system for studying early neural development. The process of early neural differentiation in hESCs begins by initiation of primitive neuroectoderm, which is manifested by rosette formation, with consecutive differentiation into neural progenitors and early glial-like cells. In this study, we examined the involvement of early neural markers - OTX2, PAX6, Sox1, Nestin, NR2F1, NR2F2, and IRX2 - in the onset of rosette formation, during spontaneous neural differentiation of hESC and human induced pluripotent stem cell (hiPSC) colonies. This is in contrast to the conventional way of studying rosette formation, which involves induction of neuronal differentiation and the utilization of embryoid bodies. Here we show that OTX2 is highly expressed at the onset of rosette formation, when rosettes comprise no more than 3-5 cells, and that its expression precedes that of established markers of early neuronal differentiation. Importantly, the rise of OTX2 expression in these cells coincides with the down-regulation of the pluripotency marker OCT4. Lastly, we show that cells derived from rosettes that emerge during spontaneous differentiation of hESCs or hiPSCs are capable of differentiating into dopaminergic neurons in vitro, and into mature-appearing pyramidal and serotonergic neurons weeks after being injected into the motor cortex of NOD-SCID mice.",

keywords = "Induced pluripotent stem cells, Mature pyramidal neurons, Neural rosettes, Radial glia",

author = "Sergey Malchenko and Jianping Xie and {de Fatima Bonaldo}, Maria and Vanin, {Elio F.} and Bhattacharyya, {Bula J.} and Abdelhak Belmadani and Guifa Xi and Vasily Galat and William Goossens and Seftor, {Richard E.B.} and Tadanori Tomita and John Crispino and Miller, {Richard J.} and Bohn, {Martha C.} and Hendrix, {Mary J.C.} and Soares, {Marcelo B.}",

note = "Funding Information: This work was supported in part by the Ann & Robert H Lurie Children's Hospital of Chicago Research Center “Excellence in Academic Medicine” Grant from the Illinois Department of Public Aid (Soares, PI) , the Dr. Ralph and Marian C. Falk Medical Research Trust , the Children Research Fund, the Gus Foundation , the Maeve McNicholas Memorial Foundation , the Everett/O'Connor Charitable Trust , NIH/NCI CA121205 (Hendrix, PI), the Medical Research Institute Council (Bohn), the Northwestern University Flow Cytometry Facility a Cancer Center Support Grant ( NCI CA060553 ), and NIH ARRA Grant RC1HL100168 (Crispino, PI). ",

year = "2014",

month = jan,

day = "25",

doi = "10.1016/j.gene.2013.07.101",

language = "English (US)",

volume = "534",

pages = "400--407",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

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Malchenko, S, Xie, J, de Fatima Bonaldo, M, Vanin, EF, Bhattacharyya, BJ, Belmadani, A, Xi, G, Galat, V, Goossens, W, Seftor, REB, Tomita, T, Crispino, J, Miller, RJ , Bohn, MC, Hendrix, MJC & Soares, MB 2014, 'Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies', Gene, vol. 534, no. 2, pp. 400-407. https://doi.org/10.1016/j.gene.2013.07.101

TY - JOUR

T1 - Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies

AU - Malchenko, Sergey

AU - Xie, Jianping

AU - de Fatima Bonaldo, Maria

AU - Vanin, Elio F.

AU - Bhattacharyya, Bula J.

AU - Belmadani, Abdelhak

AU - Xi, Guifa

AU - Galat, Vasily

AU - Goossens, William

AU - Seftor, Richard E.B.

AU - Tomita, Tadanori

AU - Crispino, John

AU - Miller, Richard J.

AU - Bohn, Martha C.

AU - Hendrix, Mary J.C.

AU - Soares, Marcelo B.

N1 - Funding Information: This work was supported in part by the Ann & Robert H Lurie Children's Hospital of Chicago Research Center “Excellence in Academic Medicine” Grant from the Illinois Department of Public Aid (Soares, PI) , the Dr. Ralph and Marian C. Falk Medical Research Trust , the Children Research Fund, the Gus Foundation , the Maeve McNicholas Memorial Foundation , the Everett/O'Connor Charitable Trust , NIH/NCI CA121205 (Hendrix, PI), the Medical Research Institute Council (Bohn), the Northwestern University Flow Cytometry Facility a Cancer Center Support Grant ( NCI CA060553 ), and NIH ARRA Grant RC1HL100168 (Crispino, PI).

PY - 2014/1/25

Y1 - 2014/1/25

N2 - In vitro neural differentiation of human embryonic stem cells (hESCs) is an advantageous system for studying early neural development. The process of early neural differentiation in hESCs begins by initiation of primitive neuroectoderm, which is manifested by rosette formation, with consecutive differentiation into neural progenitors and early glial-like cells. In this study, we examined the involvement of early neural markers - OTX2, PAX6, Sox1, Nestin, NR2F1, NR2F2, and IRX2 - in the onset of rosette formation, during spontaneous neural differentiation of hESC and human induced pluripotent stem cell (hiPSC) colonies. This is in contrast to the conventional way of studying rosette formation, which involves induction of neuronal differentiation and the utilization of embryoid bodies. Here we show that OTX2 is highly expressed at the onset of rosette formation, when rosettes comprise no more than 3-5 cells, and that its expression precedes that of established markers of early neuronal differentiation. Importantly, the rise of OTX2 expression in these cells coincides with the down-regulation of the pluripotency marker OCT4. Lastly, we show that cells derived from rosettes that emerge during spontaneous differentiation of hESCs or hiPSCs are capable of differentiating into dopaminergic neurons in vitro, and into mature-appearing pyramidal and serotonergic neurons weeks after being injected into the motor cortex of NOD-SCID mice.

AB - In vitro neural differentiation of human embryonic stem cells (hESCs) is an advantageous system for studying early neural development. The process of early neural differentiation in hESCs begins by initiation of primitive neuroectoderm, which is manifested by rosette formation, with consecutive differentiation into neural progenitors and early glial-like cells. In this study, we examined the involvement of early neural markers - OTX2, PAX6, Sox1, Nestin, NR2F1, NR2F2, and IRX2 - in the onset of rosette formation, during spontaneous neural differentiation of hESC and human induced pluripotent stem cell (hiPSC) colonies. This is in contrast to the conventional way of studying rosette formation, which involves induction of neuronal differentiation and the utilization of embryoid bodies. Here we show that OTX2 is highly expressed at the onset of rosette formation, when rosettes comprise no more than 3-5 cells, and that its expression precedes that of established markers of early neuronal differentiation. Importantly, the rise of OTX2 expression in these cells coincides with the down-regulation of the pluripotency marker OCT4. Lastly, we show that cells derived from rosettes that emerge during spontaneous differentiation of hESCs or hiPSCs are capable of differentiating into dopaminergic neurons in vitro, and into mature-appearing pyramidal and serotonergic neurons weeks after being injected into the motor cortex of NOD-SCID mice.

KW - Induced pluripotent stem cells

KW - Mature pyramidal neurons

KW - Neural rosettes

KW - Radial glia

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Onset of rosette formation during spontaneous neural differentiation of hESC and hiPSC colonies

Abstract

Funding

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