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Tumor-Free Transplantation of Patient-Derived Induced Pluripotent Stem Cell Progeny for Customized Islet Regeneration

View Article: PubMed Central - PubMed

ABSTRACT

Transplantation of progenitors from induced pluripotent stem cells reprogrammed by lentiviral vectors led to the formation of invasive teratocarcinoma-like tumors in more than 90% of immunodeficient mice. Combined transgene-free reprogramming and elimination of residual pluripotent cells by enzymatic dissociation ensured tumor-free transplantation, ultimately enabling regeneration of type 1 diabetes-specific human islet structures in vivo.

No MeSH data available.


Related in: MedlinePlus

Pancreatic progenitor cells from lentivirus-reprogrammed iPSCs (LV-iPSCs) form invasive teratocarcinoma-like tumors. (A): Stepwise differentiation protocol for generation of pancreatic endoderm cells from human iPS cells. Schematic diagram of the differentiation process is shown. (B): A representative image of left kidney, completely covered by an iPSC-derived tumor. (C): Cross-sections of the left kidney, transplanted with iPSC progeny, and the control right kidney from the same animal were imaged after H&E staining. Note the disruption of kidney structure by the iPSC-derived tumor. (D): Summary of the incidence of tumor formation upon transplantation of iPSC progeny. (E): Formation of secondary tumors upon removal of the primary tumor by unilateral nephrectomy. Secondary tumors are indicated by green arrows. (F): A metastatic tumor found on the liver surface (upper) and its cross-section (lower) are shown. Note the tumor invading deep inside the liver. (G): Immunostaining with anti-human antigen antibody was performed to detect human iPSC-derived cells in the tumors. The secondary tumors found in the peritoneal cavity, and metastatic tumors found in the lung and the liver, were shown to be human cells (shown with red signals). Nuclei were counterstained by DAPI. Abbreviations: ActA, Activin A; CYC, KAAD-cyclopamine; DAPI, 4′,6-diamidino-2-phenylindole; HPC, hematopoietic progenitor cells; ILV, indolactam V; iPSCs, induced pluripotent stem cells; ND, nondiabetic; PBMC, peripheral blood mononuclear cells; RA, all-trans retinoic acid; Wnt, Wnt 3a.
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Figure 1: Pancreatic progenitor cells from lentivirus-reprogrammed iPSCs (LV-iPSCs) form invasive teratocarcinoma-like tumors. (A): Stepwise differentiation protocol for generation of pancreatic endoderm cells from human iPS cells. Schematic diagram of the differentiation process is shown. (B): A representative image of left kidney, completely covered by an iPSC-derived tumor. (C): Cross-sections of the left kidney, transplanted with iPSC progeny, and the control right kidney from the same animal were imaged after H&E staining. Note the disruption of kidney structure by the iPSC-derived tumor. (D): Summary of the incidence of tumor formation upon transplantation of iPSC progeny. (E): Formation of secondary tumors upon removal of the primary tumor by unilateral nephrectomy. Secondary tumors are indicated by green arrows. (F): A metastatic tumor found on the liver surface (upper) and its cross-section (lower) are shown. Note the tumor invading deep inside the liver. (G): Immunostaining with anti-human antigen antibody was performed to detect human iPSC-derived cells in the tumors. The secondary tumors found in the peritoneal cavity, and metastatic tumors found in the lung and the liver, were shown to be human cells (shown with red signals). Nuclei were counterstained by DAPI. Abbreviations: ActA, Activin A; CYC, KAAD-cyclopamine; DAPI, 4′,6-diamidino-2-phenylindole; HPC, hematopoietic progenitor cells; ILV, indolactam V; iPSCs, induced pluripotent stem cells; ND, nondiabetic; PBMC, peripheral blood mononuclear cells; RA, all-trans retinoic acid; Wnt, Wnt 3a.

Mentions: Stepwise differentiation facilitates generation of PDX1- and NKX6.1-expressing pancreatic endoderm cells from pluripotent stem cell sources [22, 28]. Previous studies have demonstrated successful human islet regeneration in vivo upon transplantation of ESC-derived pancreatic endoderm/progenitor cells in immunocompromised mice [22, 29]. To assess the efficiency of human islet regeneration and the risk of teratoma formation upon transplantation of lentivirus-reprogrammed iPSC (LV-iPSC)-derived pancreatic endoderm cells, previously characterized LV-iPSCs, made from nondiabetic healthy donors and verified for their pluripotency and differentiation propensities, were differentiated into pancreatic endoderm cells (Fig. 1A), and 1 million cells were transplanted under the kidney capsule of SCID-beige mice. Within 4–6 weeks, LV-iPSC grafts gave rise to ∼2-cm solid tumors (Fig. 1B). Histology of the cross-section of the grafts demonstrated diverse cell types within the complex architecture of the graft, including glandular epithelium, adipose, muscular, and poorly differentiated tissues. Notably, the LV-iPSC-derived tumors were found invading into the kidney structure (Fig. 1C). Consistently, transplantation of pancreatic progenitors from various LV-iPSC lines resulted in rapid tumor formation in immunocompromised mice (more than 90% of cases, i.e., 17 of 18 recipient mice) (Fig. 1D).


Tumor-Free Transplantation of Patient-Derived Induced Pluripotent Stem Cell Progeny for Customized Islet Regeneration
Pancreatic progenitor cells from lentivirus-reprogrammed iPSCs (LV-iPSCs) form invasive teratocarcinoma-like tumors. (A): Stepwise differentiation protocol for generation of pancreatic endoderm cells from human iPS cells. Schematic diagram of the differentiation process is shown. (B): A representative image of left kidney, completely covered by an iPSC-derived tumor. (C): Cross-sections of the left kidney, transplanted with iPSC progeny, and the control right kidney from the same animal were imaged after H&E staining. Note the disruption of kidney structure by the iPSC-derived tumor. (D): Summary of the incidence of tumor formation upon transplantation of iPSC progeny. (E): Formation of secondary tumors upon removal of the primary tumor by unilateral nephrectomy. Secondary tumors are indicated by green arrows. (F): A metastatic tumor found on the liver surface (upper) and its cross-section (lower) are shown. Note the tumor invading deep inside the liver. (G): Immunostaining with anti-human antigen antibody was performed to detect human iPSC-derived cells in the tumors. The secondary tumors found in the peritoneal cavity, and metastatic tumors found in the lung and the liver, were shown to be human cells (shown with red signals). Nuclei were counterstained by DAPI. Abbreviations: ActA, Activin A; CYC, KAAD-cyclopamine; DAPI, 4′,6-diamidino-2-phenylindole; HPC, hematopoietic progenitor cells; ILV, indolactam V; iPSCs, induced pluripotent stem cells; ND, nondiabetic; PBMC, peripheral blood mononuclear cells; RA, all-trans retinoic acid; Wnt, Wnt 3a.
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Figure 1: Pancreatic progenitor cells from lentivirus-reprogrammed iPSCs (LV-iPSCs) form invasive teratocarcinoma-like tumors. (A): Stepwise differentiation protocol for generation of pancreatic endoderm cells from human iPS cells. Schematic diagram of the differentiation process is shown. (B): A representative image of left kidney, completely covered by an iPSC-derived tumor. (C): Cross-sections of the left kidney, transplanted with iPSC progeny, and the control right kidney from the same animal were imaged after H&E staining. Note the disruption of kidney structure by the iPSC-derived tumor. (D): Summary of the incidence of tumor formation upon transplantation of iPSC progeny. (E): Formation of secondary tumors upon removal of the primary tumor by unilateral nephrectomy. Secondary tumors are indicated by green arrows. (F): A metastatic tumor found on the liver surface (upper) and its cross-section (lower) are shown. Note the tumor invading deep inside the liver. (G): Immunostaining with anti-human antigen antibody was performed to detect human iPSC-derived cells in the tumors. The secondary tumors found in the peritoneal cavity, and metastatic tumors found in the lung and the liver, were shown to be human cells (shown with red signals). Nuclei were counterstained by DAPI. Abbreviations: ActA, Activin A; CYC, KAAD-cyclopamine; DAPI, 4′,6-diamidino-2-phenylindole; HPC, hematopoietic progenitor cells; ILV, indolactam V; iPSCs, induced pluripotent stem cells; ND, nondiabetic; PBMC, peripheral blood mononuclear cells; RA, all-trans retinoic acid; Wnt, Wnt 3a.
Mentions: Stepwise differentiation facilitates generation of PDX1- and NKX6.1-expressing pancreatic endoderm cells from pluripotent stem cell sources [22, 28]. Previous studies have demonstrated successful human islet regeneration in vivo upon transplantation of ESC-derived pancreatic endoderm/progenitor cells in immunocompromised mice [22, 29]. To assess the efficiency of human islet regeneration and the risk of teratoma formation upon transplantation of lentivirus-reprogrammed iPSC (LV-iPSC)-derived pancreatic endoderm cells, previously characterized LV-iPSCs, made from nondiabetic healthy donors and verified for their pluripotency and differentiation propensities, were differentiated into pancreatic endoderm cells (Fig. 1A), and 1 million cells were transplanted under the kidney capsule of SCID-beige mice. Within 4–6 weeks, LV-iPSC grafts gave rise to ∼2-cm solid tumors (Fig. 1B). Histology of the cross-section of the grafts demonstrated diverse cell types within the complex architecture of the graft, including glandular epithelium, adipose, muscular, and poorly differentiated tissues. Notably, the LV-iPSC-derived tumors were found invading into the kidney structure (Fig. 1C). Consistently, transplantation of pancreatic progenitors from various LV-iPSC lines resulted in rapid tumor formation in immunocompromised mice (more than 90% of cases, i.e., 17 of 18 recipient mice) (Fig. 1D).

View Article: PubMed Central - PubMed

ABSTRACT

Transplantation of progenitors from induced pluripotent stem cells reprogrammed by lentiviral vectors led to the formation of invasive teratocarcinoma-like tumors in more than 90% of immunodeficient mice. Combined transgene-free reprogramming and elimination of residual pluripotent cells by enzymatic dissociation ensured tumor-free transplantation, ultimately enabling regeneration of type 1 diabetes-specific human islet structures in vivo.

No MeSH data available.


Related in: MedlinePlus