Abstract
Purpose: The goals of this study were to optimize radiolabeling of renal lineages differentiated from human embryonic stem (hES) cells and use noninvasive imaging (positron emission tomography (PET) and bioluminescence imaging (BLI)) to detect the cells in fetal monkeys post-transplant. Procedures: hES cells expressing firefly luciferase (5×10 6) were radiolabeled with the optimized concentration of 10 μCi/ml 64Cu-PTSM then transplanted under ultrasound guidance into early second trimester fetal monkey kidneys. Fetuses were imaged in utero with PET and tissues collected for analysis 3 days post-transplant. Fetal kidneys were imaged ex vivo (PET and BLI) post-tissue harvest, and serial kidney sections were assessed by PCR for human-specific DNA sequences, fluorescent in situ hybridization (FISH) for human-specific centromere probes, and immunohistochemistry (IHC) to assess engrafted cells. Results: Transplanted cells were readily imaged in vivo and identified at the site of injection; tissue analyses confirmed the imaging findings. Using a semi-quantitative method, one in approximately 650 cells in the kidney was shown to be of human origin by PCR and FISH. Conclusions: These studies suggest that hES cells differentiated toward renal lineages can be effectively radiolabeled, transplanted into fetalmonkey kidneys under ultrasound guidance,monitored with PET post-transplant, and identified by PET, BLI, PCR, FISH, and IHC post-tissue harvest.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 197-204 |
| Number of pages | 8 |
| Journal | Molecular Imaging and Biology |
| Volume | 14 |
| Issue number | 2 |
| DOIs | |
| State | Published - Apr 2012 |
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Keywords
- Bioluminescence imaging
- Fetal transplant
- Human embryonic stem cells
- PET
- Rhesus monkey
ASJC Scopus subject areas
- Cancer Research
- Oncology
- Radiology Nuclear Medicine and imaging
Cite this
Radiolabeling and in vivo imaging of transplanted renal lineages differentiated from human embryonic stem cells in fetal rhesus monkeys. / Tarantal, Alice F; Lee, Charles C; Batchelder, Cynthia A.; Christensen, Jared E.; Prater, Daniel; Cherry, Simon R.
In: Molecular Imaging and Biology, Vol. 14, No. 2, 04.2012, p. 197-204.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Radiolabeling and in vivo imaging of transplanted renal lineages differentiated from human embryonic stem cells in fetal rhesus monkeys
AU - Tarantal, Alice F
AU - Lee, Charles C
AU - Batchelder, Cynthia A.
AU - Christensen, Jared E.
AU - Prater, Daniel
AU - Cherry, Simon R
PY - 2012/4
Y1 - 2012/4
N2 - Purpose: The goals of this study were to optimize radiolabeling of renal lineages differentiated from human embryonic stem (hES) cells and use noninvasive imaging (positron emission tomography (PET) and bioluminescence imaging (BLI)) to detect the cells in fetal monkeys post-transplant. Procedures: hES cells expressing firefly luciferase (5×10 6) were radiolabeled with the optimized concentration of 10 μCi/ml 64Cu-PTSM then transplanted under ultrasound guidance into early second trimester fetal monkey kidneys. Fetuses were imaged in utero with PET and tissues collected for analysis 3 days post-transplant. Fetal kidneys were imaged ex vivo (PET and BLI) post-tissue harvest, and serial kidney sections were assessed by PCR for human-specific DNA sequences, fluorescent in situ hybridization (FISH) for human-specific centromere probes, and immunohistochemistry (IHC) to assess engrafted cells. Results: Transplanted cells were readily imaged in vivo and identified at the site of injection; tissue analyses confirmed the imaging findings. Using a semi-quantitative method, one in approximately 650 cells in the kidney was shown to be of human origin by PCR and FISH. Conclusions: These studies suggest that hES cells differentiated toward renal lineages can be effectively radiolabeled, transplanted into fetalmonkey kidneys under ultrasound guidance,monitored with PET post-transplant, and identified by PET, BLI, PCR, FISH, and IHC post-tissue harvest.
AB - Purpose: The goals of this study were to optimize radiolabeling of renal lineages differentiated from human embryonic stem (hES) cells and use noninvasive imaging (positron emission tomography (PET) and bioluminescence imaging (BLI)) to detect the cells in fetal monkeys post-transplant. Procedures: hES cells expressing firefly luciferase (5×10 6) were radiolabeled with the optimized concentration of 10 μCi/ml 64Cu-PTSM then transplanted under ultrasound guidance into early second trimester fetal monkey kidneys. Fetuses were imaged in utero with PET and tissues collected for analysis 3 days post-transplant. Fetal kidneys were imaged ex vivo (PET and BLI) post-tissue harvest, and serial kidney sections were assessed by PCR for human-specific DNA sequences, fluorescent in situ hybridization (FISH) for human-specific centromere probes, and immunohistochemistry (IHC) to assess engrafted cells. Results: Transplanted cells were readily imaged in vivo and identified at the site of injection; tissue analyses confirmed the imaging findings. Using a semi-quantitative method, one in approximately 650 cells in the kidney was shown to be of human origin by PCR and FISH. Conclusions: These studies suggest that hES cells differentiated toward renal lineages can be effectively radiolabeled, transplanted into fetalmonkey kidneys under ultrasound guidance,monitored with PET post-transplant, and identified by PET, BLI, PCR, FISH, and IHC post-tissue harvest.
KW - Bioluminescence imaging
KW - Fetal transplant
KW - Human embryonic stem cells
KW - PET
KW - Rhesus monkey
UR - http://www.scopus.com/inward/record.url?scp=84861481756&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84861481756&partnerID=8YFLogxK
U2 - 10.1007/s11307-011-0487-1
DO - 10.1007/s11307-011-0487-1
M3 - Article
C2 - 21479709
AN - SCOPUS:84861481756
VL - 14
SP - 197
EP - 204
JO - Molecular Imaging and Biology
JF - Molecular Imaging and Biology
SN - 1536-1632
IS - 2
ER -