A scalable solution for isolating human multipotent
clinical-grade neural stem cells from ES precursors

J 2019

A scalable solution for isolating human multipotent clinical-grade neural stem cells from ES precursors

BOHAČIAKOVÁ, Dáša, Marian HRUSKA-PLOCHAN, Rachel TSUNEMOTO, Wesley D. GIFFORD, Shawn P. DRISCOLL et. al.

Basic information

Original name

A scalable solution for isolating human multipotent clinical-grade neural stem cells from ES precursors

Authors

BOHAČIAKOVÁ, Dáša (703 Slovakia, belonging to the institution), Marian HRUSKA-PLOCHAN (840 United States of America), Rachel TSUNEMOTO (840 United States of America), Wesley D. GIFFORD (840 United States of America), Shawn P. DRISCOLL (840 United States of America), Thomas D. GLENN (840 United States of America), Stephanie WU (840 United States of America), Silvia MARSALA (840 United States of America), Michael NAVARRO (840 United States of America), Takahiro TADOKORO (840 United States of America), Stefan JUHAS (203 Czech Republic), Jana JUHASOVA (203 Czech Republic), Oleksandr PLATOSHYN (840 United States of America), David PIPER (840 United States of America), Vickie SHECKLER (840 United States of America), Dara DITSWORTH (840 United States of America), Samuel L. PFAFF (840 United States of America) and Martin MARSALA (840 United States of America, guarantor)

Edition

Stem Cell Research & Therapy, London, BMC, 2019, 1757-6512

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10601 Cell biology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 5.116

RIV identification code

RIV/00216224:14110/19:00107589

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1186/s13287-019-1163-7

UT WoS

000461323900008

Keywords in English

Human embryonic stem cell (hESC); Neural stem cell (NSC); Spinal cord; Amyotrophic lateral sclerosis (ALS); Spinal traumatic injury; Bioinformatic tools to study xenografts

Abstract

V originále

Background: A well-characterized method has not yet been established to reproducibly, efficiently, and safely isolate large numbers of clinical-grade multipotent human neural stem cells (hNSCs) from embryonic stem cells (hESCs). Consequently, the transplantation of neurogenic/gliogenic precursors into the CNS for the purpose of cell replacement or neuroprotection in humans with injury or disease has not achieved widespread testing and implementation. Methods: Here, we establish an approach for the in vitro isolation of a highly expandable population of hNSCs using the manual selection of neural precursors based on their colony morphology (CoMo-NSC). The purity and NSC properties of established and extensively expanded CoMo-NSC were validated by expression of NSC markers (flow cytometry, mRNA sequencing), lack of pluripotent markers and by their tumorigenic/differentiation profile after in vivo spinal grafting in three different animal models, including (i) immunodeficient rats, (ii) immunosuppressed ALS rats (SOD1G93A), or (iii) spinally injured immunosuppressed minipigs. Results: In vitro analysis of established CoMo-NSCs showed a consistent expression of NSC markers (Sox1, Sox2, Nestin, CD24) with lack of pluripotent markers (Nanog) and stable karyotype for more than 15 passages. Gene profiling and histology revealed that spinally grafted CoMo-NSCs differentiate into neurons, astrocytes, and oligodendrocytes over a 2–6-month period in vivo without forming neoplastic derivatives or abnormal structures. Moreover, transplanted CoMo-NSCs formed neurons with synaptic contacts and glia in a variety of host environments including immunodeficient rats, immunosuppressed ALS rats (SOD1G93A), or spinally injured minipigs, indicating these cells have favorable safety and differentiation characteristics. Conclusions: These data demonstrate that manually selected CoMo-NSCs represent a safe and expandable NSC population which can effectively be used in prospective human clinical cell replacement trials for the treatment of a variety of neurodegenerative disorders, including ALS, stroke, spinal traumatic, or spinal ischemic injury.

Links

GJ15-18316Y, research and development project

Name: Úloha microRNA v řízení buněčného dělení a diferenciace lidských embryonálních kmenových buněk do neurálních kmenových buněk. (Acronym: miRNA v diferenciaci lidských EK buněk)

Investor: Czech Science Foundation

GJ18-25429Y, research and development project

Name: Funkční studie mikroRNA u nerálních kmenových buněk v průběhu diferenciace

Investor: Czech Science Foundation

Citovat

BOHAČIAKOVÁ, Dáša, Marian HRUSKA-PLOCHAN, Rachel TSUNEMOTO, Wesley D. GIFFORD, Shawn P. DRISCOLL, Thomas D. GLENN, Stephanie WU, Silvia MARSALA, Michael NAVARRO, Takahiro TADOKORO, Stefan JUHAS, Jana JUHASOVA, Oleksandr PLATOSHYN, David PIPER, Vickie SHECKLER, Dara DITSWORTH, Samuel L. PFAFF and Martin MARSALA. A scalable solution for isolating human multipotent clinical-grade neural stem cells from ES precursors. Stem Cell Research & Therapy. London: BMC, 2019, vol. 10, No 83, p. 1-19. ISSN 1757-6512. Available from: https://dx.doi.org/10.1186/s13287-019-1163-7.

@article{1550212,
   author = {Bohačiaková, Dáša and HruskaandPlochan, Marian and Tsunemoto, Rachel and Gifford, Wesley D. and Driscoll, Shawn P. and Glenn, Thomas D. and Wu, Stephanie and Marsala, Silvia and Navarro, Michael and Tadokoro, Takahiro and Juhas, Stefan and Juhasova, Jana and Platoshyn, Oleksandr and Piper, David and Sheckler, Vickie and Ditsworth, Dara and Pfaff, Samuel L. and Marsala, Martin},
   article_location = {London},
   article_number = {83},
   doi = {http://dx.doi.org/10.1186/s13287-019-1163-7},
   keywords = {Human embryonic stem cell (hESC); Neural stem cell (NSC); Spinal cord; Amyotrophic lateral sclerosis (ALS); Spinal traumatic injury; Bioinformatic tools to study xenografts},
   language = {eng},
   issn = {1757-6512},
   journal = {Stem Cell Research & Therapy},
   title = {A scalable solution for isolating human multipotent clinical-grade neural stem cells from ES precursors},
   url = {https://stemcellres.biomedcentral.com/track/pdf/10.1186/s13287-019-1163-7},
   volume = {10},
   year = {2019}
}

TY  - JOUR
ID  - 1550212
AU  - Bohačiaková, Dáša - Hruska-Plochan, Marian - Tsunemoto, Rachel - Gifford, Wesley D. - Driscoll, Shawn P. - Glenn, Thomas D. - Wu, Stephanie - Marsala, Silvia - Navarro, Michael - Tadokoro, Takahiro - Juhas, Stefan - Juhasova, Jana - Platoshyn, Oleksandr - Piper, David - Sheckler, Vickie - Ditsworth, Dara - Pfaff, Samuel L. - Marsala, Martin
PY  - 2019
TI  - A scalable solution for isolating human multipotent clinical-grade neural stem cells from ES precursors
JF  - Stem Cell Research & Therapy
VL  - 10
IS  - 83
SP  - 1-19
EP  - 1-19
PB  - BMC
SN  - 17576512
KW  - Human embryonic stem cell (hESC)
KW  - Neural stem cell (NSC)
KW  - Spinal cord
KW  - Amyotrophic lateral sclerosis (ALS)
KW  - Spinal traumatic injury
KW  - Bioinformatic tools to study xenografts
UR  - https://stemcellres.biomedcentral.com/track/pdf/10.1186/s13287-019-1163-7
L2  - https://stemcellres.biomedcentral.com/track/pdf/10.1186/s13287-019-1163-7
N2  - Background: A well-characterized method has not yet been established to reproducibly, efficiently, and safely isolate large numbers of clinical-grade multipotent human neural stem cells (hNSCs) from embryonic stem cells (hESCs). Consequently, the transplantation of neurogenic/gliogenic precursors into the CNS for the purpose of cell replacement or neuroprotection in humans with injury or disease has not achieved widespread testing and implementation. Methods: Here, we establish an approach for the in vitro isolation of a highly expandable population of hNSCs using the manual selection of neural precursors based on their colony morphology (CoMo-NSC). The purity and NSC properties of established and extensively expanded CoMo-NSC were validated by expression of NSC markers (flow cytometry, mRNA sequencing), lack of pluripotent markers and by their tumorigenic/differentiation profile after in vivo spinal grafting in three different animal models, including (i) immunodeficient rats, (ii) immunosuppressed ALS rats (SOD1G93A), or (iii) spinally injured immunosuppressed minipigs. Results: In vitro analysis of established CoMo-NSCs showed a consistent expression of NSC markers (Sox1, Sox2, Nestin, CD24) with lack of pluripotent markers (Nanog) and stable karyotype for more than 15 passages. Gene profiling and histology revealed that spinally grafted CoMo-NSCs differentiate into neurons, astrocytes, and oligodendrocytes over a 2–6-month period in vivo without forming neoplastic derivatives or abnormal structures. Moreover, transplanted CoMo-NSCs formed neurons with synaptic contacts and glia in a variety of host environments including immunodeficient rats, immunosuppressed ALS rats (SOD1G93A), or spinally injured minipigs, indicating these cells have favorable safety and differentiation characteristics. Conclusions: These data demonstrate that manually selected CoMo-NSCs represent a safe and expandable NSC population which can effectively be used in prospective human clinical cell replacement trials for the treatment of a variety of neurodegenerative disorders, including ALS, stroke, spinal traumatic, or spinal ischemic injury.
ER  -

BOHAČIAKOVÁ, Dáša, Marian HRUSKA-PLOCHAN, Rachel TSUNEMOTO, Wesley D. GIFFORD, Shawn P. DRISCOLL, Thomas D. GLENN, Stephanie WU, Silvia MARSALA, Michael NAVARRO, Takahiro TADOKORO, Stefan JUHAS, Jana JUHASOVA, Oleksandr PLATOSHYN, David PIPER, Vickie SHECKLER, Dara DITSWORTH, Samuel L. PFAFF and Martin MARSALA. A scalable solution for isolating human multipotent clinical-grade neural stem cells from ES precursors. \textit{Stem Cell Research \&{} Therapy}. London: BMC, 2019, vol.~10, No~83, p.~1-19. ISSN~1757-6512. Available from: https://dx.doi.org/10.1186/s13287-019-1163-7.

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