Real-time in vivo imaging of stem cells following transgenesis by transposition

Jakub Tolar; Mark Osborn; Scott Bell; Ron McElmurry; Lily Xia; Megan Riddle; Angela Panoskaltsis-Mortari; Yuehua Jiang; R. Scott McIvor; Christopher H. Contag; Stephen R. Yant; Mark A. Kay; Catherine M. Verfaillie; Bruce R. Blazar

doi:10.1016/j.ymthe.2005.02.023

Real-time in vivo imaging of stem cells following transgenesis by transposition

Jakub Tolar, Mark Osborn, Scott Bell, Ron McElmurry, Lily Xia, Megan Riddle, Angela Panoskaltsis-Mortari, Yuehua Jiang, R. Scott McIvor, Christopher H. Contag, Stephen R. Yant, Mark A. Kay, Catherine M. Verfaillie, Bruce R. Blazar

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

34 Scopus citations

Abstract

Previous studies have identified Sleeping Beauty transposons as efficient vectors for nonviral gene delivery in mammalian cells. However, studies demonstrating the usefulness of transposons as gene delivery vehicles into adult stem cells are lacking. Multipotent adult progenitor cells (MAPC) are nonhematopoietic stem cells with the capacity to form most, if not all, cell types of the body and as such hold great therapeutic potential. The whole-body biodistribution and persistence of MAPC are unknown, and such data would help direct clinical applications. We have nucleofected murine MAPC with two plasmid-based Sleeping Beauty transposons encoding the red fluorescent protein (DsRed2) and firefly luciferase. Transgenic euploid MAPC clones maintained their characteristic multilineage differentiation potential in vitro. DsRed2 and luciferase expression allowed for MAPC detection in vivo and in tissue sections. To confirm that transgenesis occurred by transposition into the genome of MAPC, we mapped Sleeping Beauty transposon integration sites in two MAPC clones using splinkerette PCR. This novel dual-reporter imaging approach based on the transgenesis of MAPC with Sleeping Beauty transposons sheds light on the homing patterns of MAPC and paves the way for quantification of MAPC engraftment in real time in vivo.

Original language	English (US)
Pages (from-to)	42-48
Number of pages	7
Journal	Molecular Therapy
Volume	12
Issue number	1
DOIs	https://doi.org/10.1016/j.ymthe.2005.02.023
State	Published - Jul 2005

Bibliographical note

Funding Information:
This work was supported in part by the ASCO Young Investigator Award, the Fanconi Anemia Research Fund, the Children’s Cancer Research Fund, NIH Grants RO1 HL55209, R01 HL49997, R01 HL52952, and R24 CA 92862 (CHC), and the Child Health Research Scholar Award (NIH 5K12-HD033692-10). The confocal microscope was purchased using NCRR shared instrumentation Grant 1 S10 RR16851. We thank Sara Benning, Kevin Tram, and Amy Nordlander for their technical assistance.

Keywords

Biophotonic imaging
DsRed2
Luciferase
Multipotent adult progenitor cell
Sleeping Beauty
Stem cell homing
Whole-body imaging

Access

10.1016/j.ymthe.2005.02.023

OpenUrl availability

Full text

Cite this

@article{62e20684b4614b258e28f1fe520c1e54,

title = "Real-time in vivo imaging of stem cells following transgenesis by transposition",

abstract = "Previous studies have identified Sleeping Beauty transposons as efficient vectors for nonviral gene delivery in mammalian cells. However, studies demonstrating the usefulness of transposons as gene delivery vehicles into adult stem cells are lacking. Multipotent adult progenitor cells (MAPC) are nonhematopoietic stem cells with the capacity to form most, if not all, cell types of the body and as such hold great therapeutic potential. The whole-body biodistribution and persistence of MAPC are unknown, and such data would help direct clinical applications. We have nucleofected murine MAPC with two plasmid-based Sleeping Beauty transposons encoding the red fluorescent protein (DsRed2) and firefly luciferase. Transgenic euploid MAPC clones maintained their characteristic multilineage differentiation potential in vitro. DsRed2 and luciferase expression allowed for MAPC detection in vivo and in tissue sections. To confirm that transgenesis occurred by transposition into the genome of MAPC, we mapped Sleeping Beauty transposon integration sites in two MAPC clones using splinkerette PCR. This novel dual-reporter imaging approach based on the transgenesis of MAPC with Sleeping Beauty transposons sheds light on the homing patterns of MAPC and paves the way for quantification of MAPC engraftment in real time in vivo.",

keywords = "Biophotonic imaging, DsRed2, Luciferase, Multipotent adult progenitor cell, Sleeping Beauty, Stem cell homing, Whole-body imaging",

author = "Jakub Tolar and Mark Osborn and Scott Bell and Ron McElmurry and Lily Xia and Megan Riddle and Angela Panoskaltsis-Mortari and Yuehua Jiang and McIvor, {R. Scott} and Contag, {Christopher H.} and Yant, {Stephen R.} and Kay, {Mark A.} and Verfaillie, {Catherine M.} and Blazar, {Bruce R.}",

note = "Funding Information: This work was supported in part by the ASCO Young Investigator Award, the Fanconi Anemia Research Fund, the Children{\textquoteright}s Cancer Research Fund, NIH Grants RO1 HL55209, R01 HL49997, R01 HL52952, and R24 CA 92862 (CHC), and the Child Health Research Scholar Award (NIH 5K12-HD033692-10). The confocal microscope was purchased using NCRR shared instrumentation Grant 1 S10 RR16851. We thank Sara Benning, Kevin Tram, and Amy Nordlander for their technical assistance.",

year = "2005",

month = jul,

doi = "10.1016/j.ymthe.2005.02.023",

language = "English (US)",

volume = "12",

pages = "42--48",

journal = "Molecular Therapy",

issn = "1525-0016",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Real-time in vivo imaging of stem cells following transgenesis by transposition

AU - Tolar, Jakub

AU - Osborn, Mark

AU - Bell, Scott

AU - McElmurry, Ron

AU - Xia, Lily

AU - Riddle, Megan

AU - Panoskaltsis-Mortari, Angela

AU - Jiang, Yuehua

AU - McIvor, R. Scott

AU - Contag, Christopher H.

AU - Yant, Stephen R.

AU - Kay, Mark A.

AU - Verfaillie, Catherine M.

AU - Blazar, Bruce R.

N1 - Funding Information: This work was supported in part by the ASCO Young Investigator Award, the Fanconi Anemia Research Fund, the Children’s Cancer Research Fund, NIH Grants RO1 HL55209, R01 HL49997, R01 HL52952, and R24 CA 92862 (CHC), and the Child Health Research Scholar Award (NIH 5K12-HD033692-10). The confocal microscope was purchased using NCRR shared instrumentation Grant 1 S10 RR16851. We thank Sara Benning, Kevin Tram, and Amy Nordlander for their technical assistance.

PY - 2005/7

Y1 - 2005/7

N2 - Previous studies have identified Sleeping Beauty transposons as efficient vectors for nonviral gene delivery in mammalian cells. However, studies demonstrating the usefulness of transposons as gene delivery vehicles into adult stem cells are lacking. Multipotent adult progenitor cells (MAPC) are nonhematopoietic stem cells with the capacity to form most, if not all, cell types of the body and as such hold great therapeutic potential. The whole-body biodistribution and persistence of MAPC are unknown, and such data would help direct clinical applications. We have nucleofected murine MAPC with two plasmid-based Sleeping Beauty transposons encoding the red fluorescent protein (DsRed2) and firefly luciferase. Transgenic euploid MAPC clones maintained their characteristic multilineage differentiation potential in vitro. DsRed2 and luciferase expression allowed for MAPC detection in vivo and in tissue sections. To confirm that transgenesis occurred by transposition into the genome of MAPC, we mapped Sleeping Beauty transposon integration sites in two MAPC clones using splinkerette PCR. This novel dual-reporter imaging approach based on the transgenesis of MAPC with Sleeping Beauty transposons sheds light on the homing patterns of MAPC and paves the way for quantification of MAPC engraftment in real time in vivo.

AB - Previous studies have identified Sleeping Beauty transposons as efficient vectors for nonviral gene delivery in mammalian cells. However, studies demonstrating the usefulness of transposons as gene delivery vehicles into adult stem cells are lacking. Multipotent adult progenitor cells (MAPC) are nonhematopoietic stem cells with the capacity to form most, if not all, cell types of the body and as such hold great therapeutic potential. The whole-body biodistribution and persistence of MAPC are unknown, and such data would help direct clinical applications. We have nucleofected murine MAPC with two plasmid-based Sleeping Beauty transposons encoding the red fluorescent protein (DsRed2) and firefly luciferase. Transgenic euploid MAPC clones maintained their characteristic multilineage differentiation potential in vitro. DsRed2 and luciferase expression allowed for MAPC detection in vivo and in tissue sections. To confirm that transgenesis occurred by transposition into the genome of MAPC, we mapped Sleeping Beauty transposon integration sites in two MAPC clones using splinkerette PCR. This novel dual-reporter imaging approach based on the transgenesis of MAPC with Sleeping Beauty transposons sheds light on the homing patterns of MAPC and paves the way for quantification of MAPC engraftment in real time in vivo.

KW - Biophotonic imaging

KW - DsRed2

KW - Luciferase

KW - Multipotent adult progenitor cell

KW - Sleeping Beauty

KW - Stem cell homing

KW - Whole-body imaging

UR - http://www.scopus.com/inward/record.url?scp=20844447232&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20844447232&partnerID=8YFLogxK

U2 - 10.1016/j.ymthe.2005.02.023

DO - 10.1016/j.ymthe.2005.02.023

M3 - Article

C2 - 15963919

AN - SCOPUS:20844447232

SN - 1525-0016

VL - 12

SP - 42

EP - 48

JO - Molecular Therapy

JF - Molecular Therapy

IS - 1

ER -

Real-time in vivo imaging of stem cells following transgenesis by transposition

Abstract

Bibliographical note

Keywords

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this