Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e

Talmadge, M. Hirai, D. Laface, S. N. Robinson, L. Kelsey, R. Johnson, S. Fen Wen, Phyllis Irene Warkentin, K. Mills, M. Vaillancourt, J. Chavez, C. Leutzinger, J. Sumegi, S. Neugebauer, J. Lehman, O. Talmadge, D. C. Maneval

Research output: Contribution to journalArticle

Abstract

Replication-deficient, recombinant adenovirus carrying the wild-type p53 gene (rAdp53) can significantly reduce tumor cell contamination of stem cell products from bre ist cancer patients. This provides a useful tumor cell purging strategy, which we have previous ly shown to have six log of activity. However, the safety of this procedure for self-renewii g, pluripotent hematopoietic stem cells has not been determined. The ability of human st(m cells to reconstitute hematopoiesis in mice with severe combined immunodeficiency (SCI D) and to undergo secondary transplantation provides a measure of self-renewing stem c ;11 activity. In these studies we examined the effect of rAd-p53 co-incubation with stem c;ll products on SCID «populating activity (SRA). At the targeted clinical dose (2 x I')10 particles/ml), rAd-p53 was co-cultured with mobilized human CD34 cells. The ability of these cells to establish multi-lineage, human-derived hematopoiesis in sublethally irradiat< :d, non-obese diabetic (NOD)-SCID mice was investigated. The presence of human cells w as determined by flow cytometry, in vitro granulocyte-macrophage colony-forming u lit assay and the demonstration of human Alu-sequences by PCR and Southern blottiiig. Limiting dilution analysis (LDA) provided a quantitative comparison between the SRA of control CD34 cells and CD34 cells co-incubated with rAd-p53. The LDA results in 1 le NOD-SCID mouse 6 weeks after transplantation were 1:94,170 (95% CI: 56,391 -157,256) and 1:50,747 (95% CI: 29,662 - 86,819) in the rAd-p53 co-incubated and control CD; 4' samples, respectively. Similar results were observed using flow cytometry data. Six wee ks after transplantation, marrow cells were harvested from the primary recipients and transplanted into secondary NOD-SCID mice. After 2 weeks, human cells were found in the marrow and spleen by flow cytometry. We conclude that the co-incubation of stem cell products with rAd-p53 is a safe purging protocol with little effect on self-renewiijg, pluripotent stem cells.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART I
StatePublished - Dec 1 2000

Fingerprint

p53 Genes
Stem cells
Genes
Cells
Flow cytometry
Purging
Inbred NOD Mouse
SCID Mice
Dilution
Tumors
Pluripotent Stem Cells
Flow Cytometry
Hematopoiesis
Macrophages
Stem Cells
Transplantation
Assays
Bone Marrow
Contamination
Demonstrations

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Talmadge, Hirai, M., Laface, D., Robinson, S. N., Kelsey, L., Johnson, R., ... Maneval, D. C. (2000). Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e. Blood, 96(11 PART I).

Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e. / Talmadge; Hirai, M.; Laface, D.; Robinson, S. N.; Kelsey, L.; Johnson, R.; Fen Wen, S.; Warkentin, Phyllis Irene; Mills, K.; Vaillancourt, M.; Chavez, J.; Leutzinger, C.; Sumegi, J.; Neugebauer, S.; Lehman, J.; Talmadge, O.; Maneval, D. C.

In: Blood, Vol. 96, No. 11 PART I, 01.12.2000.

Research output: Contribution to journalArticle

Talmadge, Hirai, M, Laface, D, Robinson, SN, Kelsey, L, Johnson, R, Fen Wen, S, Warkentin, PI, Mills, K, Vaillancourt, M, Chavez, J, Leutzinger, C, Sumegi, J, Neugebauer, S, Lehman, J, Talmadge, O & Maneval, DC 2000, 'Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e', Blood, vol. 96, no. 11 PART I.
Talmadge ; Hirai, M. ; Laface, D. ; Robinson, S. N. ; Kelsey, L. ; Johnson, R. ; Fen Wen, S. ; Warkentin, Phyllis Irene ; Mills, K. ; Vaillancourt, M. ; Chavez, J. ; Leutzinger, C. ; Sumegi, J. ; Neugebauer, S. ; Lehman, J. ; Talmadge, O. ; Maneval, D. C. / Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e. In: Blood. 2000 ; Vol. 96, No. 11 PART I.
@article{39c1ccecc79747e996b4586d49569859,
title = "Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e",
abstract = "Replication-deficient, recombinant adenovirus carrying the wild-type p53 gene (rAdp53) can significantly reduce tumor cell contamination of stem cell products from bre ist cancer patients. This provides a useful tumor cell purging strategy, which we have previous ly shown to have six log of activity. However, the safety of this procedure for self-renewii g, pluripotent hematopoietic stem cells has not been determined. The ability of human st(m cells to reconstitute hematopoiesis in mice with severe combined immunodeficiency (SCI D) and to undergo secondary transplantation provides a measure of self-renewing stem c ;11 activity. In these studies we examined the effect of rAd-p53 co-incubation with stem c;ll products on SCID «populating activity (SRA). At the targeted clinical dose (2 x I')10 particles/ml), rAd-p53 was co-cultured with mobilized human CD34 cells. The ability of these cells to establish multi-lineage, human-derived hematopoiesis in sublethally irradiat< :d, non-obese diabetic (NOD)-SCID mice was investigated. The presence of human cells w as determined by flow cytometry, in vitro granulocyte-macrophage colony-forming u lit assay and the demonstration of human Alu-sequences by PCR and Southern blottiiig. Limiting dilution analysis (LDA) provided a quantitative comparison between the SRA of control CD34 cells and CD34 cells co-incubated with rAd-p53. The LDA results in 1 le NOD-SCID mouse 6 weeks after transplantation were 1:94,170 (95{\%} CI: 56,391 -157,256) and 1:50,747 (95{\%} CI: 29,662 - 86,819) in the rAd-p53 co-incubated and control CD; 4' samples, respectively. Similar results were observed using flow cytometry data. Six wee ks after transplantation, marrow cells were harvested from the primary recipients and transplanted into secondary NOD-SCID mice. After 2 weeks, human cells were found in the marrow and spleen by flow cytometry. We conclude that the co-incubation of stem cell products with rAd-p53 is a safe purging protocol with little effect on self-renewiijg, pluripotent stem cells.",
author = "Talmadge and M. Hirai and D. Laface and Robinson, {S. N.} and L. Kelsey and R. Johnson and {Fen Wen}, S. and Warkentin, {Phyllis Irene} and K. Mills and M. Vaillancourt and J. Chavez and C. Leutzinger and J. Sumegi and S. Neugebauer and J. Lehman and O. Talmadge and Maneval, {D. C.}",
year = "2000",
month = "12",
day = "1",
language = "English (US)",
volume = "96",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "11 PART I",

}

TY - JOUR

T1 - Nod-scid reconstitution by human cd34+ cells treatead with a replication-deficient, recombinant adenovirl fs vector containing the wild-type p53 gene (rad-p53). J. e

AU - Talmadge,

AU - Hirai, M.

AU - Laface, D.

AU - Robinson, S. N.

AU - Kelsey, L.

AU - Johnson, R.

AU - Fen Wen, S.

AU - Warkentin, Phyllis Irene

AU - Mills, K.

AU - Vaillancourt, M.

AU - Chavez, J.

AU - Leutzinger, C.

AU - Sumegi, J.

AU - Neugebauer, S.

AU - Lehman, J.

AU - Talmadge, O.

AU - Maneval, D. C.

PY - 2000/12/1

Y1 - 2000/12/1

N2 - Replication-deficient, recombinant adenovirus carrying the wild-type p53 gene (rAdp53) can significantly reduce tumor cell contamination of stem cell products from bre ist cancer patients. This provides a useful tumor cell purging strategy, which we have previous ly shown to have six log of activity. However, the safety of this procedure for self-renewii g, pluripotent hematopoietic stem cells has not been determined. The ability of human st(m cells to reconstitute hematopoiesis in mice with severe combined immunodeficiency (SCI D) and to undergo secondary transplantation provides a measure of self-renewing stem c ;11 activity. In these studies we examined the effect of rAd-p53 co-incubation with stem c;ll products on SCID «populating activity (SRA). At the targeted clinical dose (2 x I')10 particles/ml), rAd-p53 was co-cultured with mobilized human CD34 cells. The ability of these cells to establish multi-lineage, human-derived hematopoiesis in sublethally irradiat< :d, non-obese diabetic (NOD)-SCID mice was investigated. The presence of human cells w as determined by flow cytometry, in vitro granulocyte-macrophage colony-forming u lit assay and the demonstration of human Alu-sequences by PCR and Southern blottiiig. Limiting dilution analysis (LDA) provided a quantitative comparison between the SRA of control CD34 cells and CD34 cells co-incubated with rAd-p53. The LDA results in 1 le NOD-SCID mouse 6 weeks after transplantation were 1:94,170 (95% CI: 56,391 -157,256) and 1:50,747 (95% CI: 29,662 - 86,819) in the rAd-p53 co-incubated and control CD; 4' samples, respectively. Similar results were observed using flow cytometry data. Six wee ks after transplantation, marrow cells were harvested from the primary recipients and transplanted into secondary NOD-SCID mice. After 2 weeks, human cells were found in the marrow and spleen by flow cytometry. We conclude that the co-incubation of stem cell products with rAd-p53 is a safe purging protocol with little effect on self-renewiijg, pluripotent stem cells.

AB - Replication-deficient, recombinant adenovirus carrying the wild-type p53 gene (rAdp53) can significantly reduce tumor cell contamination of stem cell products from bre ist cancer patients. This provides a useful tumor cell purging strategy, which we have previous ly shown to have six log of activity. However, the safety of this procedure for self-renewii g, pluripotent hematopoietic stem cells has not been determined. The ability of human st(m cells to reconstitute hematopoiesis in mice with severe combined immunodeficiency (SCI D) and to undergo secondary transplantation provides a measure of self-renewing stem c ;11 activity. In these studies we examined the effect of rAd-p53 co-incubation with stem c;ll products on SCID «populating activity (SRA). At the targeted clinical dose (2 x I')10 particles/ml), rAd-p53 was co-cultured with mobilized human CD34 cells. The ability of these cells to establish multi-lineage, human-derived hematopoiesis in sublethally irradiat< :d, non-obese diabetic (NOD)-SCID mice was investigated. The presence of human cells w as determined by flow cytometry, in vitro granulocyte-macrophage colony-forming u lit assay and the demonstration of human Alu-sequences by PCR and Southern blottiiig. Limiting dilution analysis (LDA) provided a quantitative comparison between the SRA of control CD34 cells and CD34 cells co-incubated with rAd-p53. The LDA results in 1 le NOD-SCID mouse 6 weeks after transplantation were 1:94,170 (95% CI: 56,391 -157,256) and 1:50,747 (95% CI: 29,662 - 86,819) in the rAd-p53 co-incubated and control CD; 4' samples, respectively. Similar results were observed using flow cytometry data. Six wee ks after transplantation, marrow cells were harvested from the primary recipients and transplanted into secondary NOD-SCID mice. After 2 weeks, human cells were found in the marrow and spleen by flow cytometry. We conclude that the co-incubation of stem cell products with rAd-p53 is a safe purging protocol with little effect on self-renewiijg, pluripotent stem cells.

UR - http://www.scopus.com/inward/record.url?scp=33748654396&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748654396&partnerID=8YFLogxK

M3 - Article

VL - 96

JO - Blood

JF - Blood

SN - 0006-4971

IS - 11 PART I

ER -