Directed engineering of umbilical cord blood stem cells to produce C-peptide and insulin

Larry Denner, Y. Bodenburg, J. G. Zhao, M. Howe, J. Cappo, Ronald Tilton, J. A. Copland, N. Forraz, C. McGuckin, Randall Urban

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Objectives: In this study, we investigated the potential of umbilical cord blood stem cell lineages to produce C-peptide and insulin. Materials and methods: Lineage negative, CD133+ and CD34+ cells were analyzed by flow cytometry to assess expression of cell division antigens. These lineages were expanded in culture and subjected to an established protocol to differentiate mouse embryonic stem cells (ESCs) toward the pancreatic phenotype. Phase contrast and fluorescence immunocytochemistry were used to characterize differentiation markers with particular emphasis on insulin and C-peptide. Results: All 3 lineages expressed SSEA-4, a marker previously reported to be restricted to the ESC compartment. Phase contrast microscopy showed all three lineages recapitulated the treatment-dependent morphological changes of ESCs as well as the temporally restricted expression of nestin and vimentin during differentiation. After engineering, each isolate contained both C-peptide and insulin, a result also obtained following a much shorter protocol for ESCs. Conclusions: Since C-peptide can only be derived from de novo synthesis and processing of pre-proinsulin mRNA and protein, we conclude that these results are the first demonstration that human umbilical cord blood-derived stem cells can be engineered to engage in de novo synthesis of insulin.

Original languageEnglish (US)
Pages (from-to)367-380
Number of pages14
JournalCell Proliferation
Volume40
Issue number3
DOIs
StatePublished - Jun 2007

Fingerprint

C-Peptide
Fetal Blood
Blood Cells
Stem Cells
Embryonic Stem Cells
Insulin
Phase-Contrast Microscopy
Nestin
RNA Precursors
Differentiation Antigens
Vimentin
Cell Lineage
Cell Division
Flow Cytometry
Fluorescence
Immunohistochemistry
Phenotype
Antigens
Proteins
Therapeutics

ASJC Scopus subject areas

  • Cell Biology

Cite this

Directed engineering of umbilical cord blood stem cells to produce C-peptide and insulin. / Denner, Larry; Bodenburg, Y.; Zhao, J. G.; Howe, M.; Cappo, J.; Tilton, Ronald; Copland, J. A.; Forraz, N.; McGuckin, C.; Urban, Randall.

In: Cell Proliferation, Vol. 40, No. 3, 06.2007, p. 367-380.

Research output: Contribution to journalArticle

Denner, L, Bodenburg, Y, Zhao, JG, Howe, M, Cappo, J, Tilton, R, Copland, JA, Forraz, N, McGuckin, C & Urban, R 2007, 'Directed engineering of umbilical cord blood stem cells to produce C-peptide and insulin', Cell Proliferation, vol. 40, no. 3, pp. 367-380. https://doi.org/10.1111/j.1365-2184.2007.00439.x
Denner, Larry ; Bodenburg, Y. ; Zhao, J. G. ; Howe, M. ; Cappo, J. ; Tilton, Ronald ; Copland, J. A. ; Forraz, N. ; McGuckin, C. ; Urban, Randall. / Directed engineering of umbilical cord blood stem cells to produce C-peptide and insulin. In: Cell Proliferation. 2007 ; Vol. 40, No. 3. pp. 367-380.
@article{63d9c56ee04e4fb2838f67c7de54769c,
title = "Directed engineering of umbilical cord blood stem cells to produce C-peptide and insulin",
abstract = "Objectives: In this study, we investigated the potential of umbilical cord blood stem cell lineages to produce C-peptide and insulin. Materials and methods: Lineage negative, CD133+ and CD34+ cells were analyzed by flow cytometry to assess expression of cell division antigens. These lineages were expanded in culture and subjected to an established protocol to differentiate mouse embryonic stem cells (ESCs) toward the pancreatic phenotype. Phase contrast and fluorescence immunocytochemistry were used to characterize differentiation markers with particular emphasis on insulin and C-peptide. Results: All 3 lineages expressed SSEA-4, a marker previously reported to be restricted to the ESC compartment. Phase contrast microscopy showed all three lineages recapitulated the treatment-dependent morphological changes of ESCs as well as the temporally restricted expression of nestin and vimentin during differentiation. After engineering, each isolate contained both C-peptide and insulin, a result also obtained following a much shorter protocol for ESCs. Conclusions: Since C-peptide can only be derived from de novo synthesis and processing of pre-proinsulin mRNA and protein, we conclude that these results are the first demonstration that human umbilical cord blood-derived stem cells can be engineered to engage in de novo synthesis of insulin.",
author = "Larry Denner and Y. Bodenburg and Zhao, {J. G.} and M. Howe and J. Cappo and Ronald Tilton and Copland, {J. A.} and N. Forraz and C. McGuckin and Randall Urban",
year = "2007",
month = "6",
doi = "10.1111/j.1365-2184.2007.00439.x",
language = "English (US)",
volume = "40",
pages = "367--380",
journal = "Cell Proliferation",
issn = "0960-7722",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Directed engineering of umbilical cord blood stem cells to produce C-peptide and insulin

AU - Denner, Larry

AU - Bodenburg, Y.

AU - Zhao, J. G.

AU - Howe, M.

AU - Cappo, J.

AU - Tilton, Ronald

AU - Copland, J. A.

AU - Forraz, N.

AU - McGuckin, C.

AU - Urban, Randall

PY - 2007/6

Y1 - 2007/6

N2 - Objectives: In this study, we investigated the potential of umbilical cord blood stem cell lineages to produce C-peptide and insulin. Materials and methods: Lineage negative, CD133+ and CD34+ cells were analyzed by flow cytometry to assess expression of cell division antigens. These lineages were expanded in culture and subjected to an established protocol to differentiate mouse embryonic stem cells (ESCs) toward the pancreatic phenotype. Phase contrast and fluorescence immunocytochemistry were used to characterize differentiation markers with particular emphasis on insulin and C-peptide. Results: All 3 lineages expressed SSEA-4, a marker previously reported to be restricted to the ESC compartment. Phase contrast microscopy showed all three lineages recapitulated the treatment-dependent morphological changes of ESCs as well as the temporally restricted expression of nestin and vimentin during differentiation. After engineering, each isolate contained both C-peptide and insulin, a result also obtained following a much shorter protocol for ESCs. Conclusions: Since C-peptide can only be derived from de novo synthesis and processing of pre-proinsulin mRNA and protein, we conclude that these results are the first demonstration that human umbilical cord blood-derived stem cells can be engineered to engage in de novo synthesis of insulin.

AB - Objectives: In this study, we investigated the potential of umbilical cord blood stem cell lineages to produce C-peptide and insulin. Materials and methods: Lineage negative, CD133+ and CD34+ cells were analyzed by flow cytometry to assess expression of cell division antigens. These lineages were expanded in culture and subjected to an established protocol to differentiate mouse embryonic stem cells (ESCs) toward the pancreatic phenotype. Phase contrast and fluorescence immunocytochemistry were used to characterize differentiation markers with particular emphasis on insulin and C-peptide. Results: All 3 lineages expressed SSEA-4, a marker previously reported to be restricted to the ESC compartment. Phase contrast microscopy showed all three lineages recapitulated the treatment-dependent morphological changes of ESCs as well as the temporally restricted expression of nestin and vimentin during differentiation. After engineering, each isolate contained both C-peptide and insulin, a result also obtained following a much shorter protocol for ESCs. Conclusions: Since C-peptide can only be derived from de novo synthesis and processing of pre-proinsulin mRNA and protein, we conclude that these results are the first demonstration that human umbilical cord blood-derived stem cells can be engineered to engage in de novo synthesis of insulin.

UR - http://www.scopus.com/inward/record.url?scp=34249019166&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34249019166&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2184.2007.00439.x

DO - 10.1111/j.1365-2184.2007.00439.x

M3 - Article

VL - 40

SP - 367

EP - 380

JO - Cell Proliferation

JF - Cell Proliferation

SN - 0960-7722

IS - 3

ER -