In vivo generation of beta-cell-like cells from CD34(+) cells differentiated from human embryonic stem cells.
OBJECTIVE
CD34(+) cells, present within the bone marrow, have previously been shown to possess pancreatic endocrine potential. Based on this observation, we explored the capacity of CD34(+) cells derived in culture from the differentiation of human embryonic stem cells (hESC), for their in vivo pancreatic endocrine capacity.


METHODS
Sheep were transplanted with hESC-derived CD34(+) cells, as well as nonsorted differentiated cultures. Transplantations were carried out with in utero intraperitoneal injections prior to development of the immune system in the fetus so that tolerance toward foreign antigens was acquired during gestation and persisted in the adult.


RESULTS
All cell populations that were tested demonstrated human cellular activity and long-term presence up to 5 years. However, the in vivo beta-cell-like activity achieved from the transplantation of the sorted CD34(+) cell population was not augmented by transplanting the entire cell population from which the CD34(+) cells were isolated. Human DNA and insulin messenger RNA were detected in sheep pancreases. An average of 1.51 ng/mL human C-peptide was detected in serum from eight animals transplanted with differentiated cell populations and assayed up to 55 months posttransplantation. Transplantation of as few as 23,500 cells resulted in long-term sustainable beta-cell-like activity. Teratomas were absent in the transplanted animals.


CONCLUSIONS
Our data suggest that hESC-derived CD34(+) cells have a potential for long-term in vivo endocrine cellular activity that could prove useful in regenerative medicine. Because the same cell population has previously been shown to contain hematopoietic potential, it could be used for the induction of immunological tolerance and bone marrow chimerism prior to cellular therapy for diabetes.
Goodrich
A Daisy
AD
Department of Animal Biotechnology, University of Nevada, Reno, Reno, NV 89557, USA. daisy@med.unr.edu
Ersek
Adel
A
Varain
Nicole M
NM
Groza
Daria
D
Cenariu
Mihai
M
Thain
David S
DS
Almeida-Porada
Graca
G
Porada
Christopher D
CD
Zanjani
Esmail D
ED
D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology*; D001483:Base Sequence; D001786:Blood Glucose / Q000032:analysis; D002454:Cell Differentiation; D004247:DNA / Q000032:analysis; D017931:DNA Primers; D053595:Embryonic Stem Cells / Q000166:cytology / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000235:genetics; D007515:Islets of Langerhans / Q000166:cytology* / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep
D018952:Antigens, CD34; D001786:Blood Glucose; D017931:DNA Primers; D007328:Insulin; D012333:RNA, Messenger; D004247:DNA