How old is too old?
Colony-forming units assay
Cryopreservation
Hematopoietic stem cell transplantation
Hematopoietic stem cells
In vitro techniques
Long-term storage
Peripheral blood stem cell
Transplant
Viability
Journal
World journal of stem cells
ISSN: 1948-0210
Titre abrégé: World J Stem Cells
Pays: United States
ID NLM: 101535826
Informations de publication
Date de publication:
26 May 2020
26 May 2020
Historique:
received:
28
02
2020
revised:
14
04
2020
accepted:
28
04
2020
entrez:
18
6
2020
pubmed:
18
6
2020
medline:
18
6
2020
Statut:
ppublish
Résumé
Peripheral blood stem cells (PBSC) are commonly cryopreserved awaiting clinical use for hematopoietic stem cell transplant. Long term cryopreservation is commonly defined as five years or longer, and limited data exists regarding how long PBSC can be cryopreserved and retain the ability to successfully engraft. Clinical programs, stem cell banks, and regulatory and accrediting agencies interested in product stability would benefit from such data. Thus, we assessed recovery and colony forming ability of PBSC following long-term cryopreservation as well as their ability to engraft in NOD/SCID/IL-2Rγ To investigate the in vivo engraftment potential of long-term cryopreserved PBSC units. PBSC units which were collected and frozen using validated clinical protocols were obtained for research use from the Cellular Therapy Laboratory at Indiana University Health. These units were thawed in the Cellular Therapy Laboratory using clinical standards of practice, and the pre-freeze and post-thaw characteristics of the units were compared. Progenitor function was assessed using standard colony-forming assays. CD34-selected cells were transplanted into immunodeficient mice to assess stem cell function. Ten PBSC units with mean of 17 years in cryopreservation (range 13.6-18.3 years) demonstrated a mean total cell recovery of 88% ± 12% (range 68%-110%) and post-thaw viability of 69% ± 17% (range 34%-86%). BFU-E growth was shown in 9 of 10 units and CFU-GM growth in 7 of 10 units post-thaw. Immunodeficient mice were transplanted with CD34-selected cells from four randomly chosen PBSC units. All mice demonstrated long-term engraftment at 12 wk with mean 34% ± 24% human CD45+ cells, and differentiation with presence of human CD19+, CD3+ and CD33+ cells. Harvested bone marrow from all mice demonstrated growth of erythroid and myeloid colonies. We demonstrated engraftment of clinically-collected and thawed PBSC following cryopreservation up to 18 years in NSG mice, signifying likely successful clinical transplantation of PBSC following long-term cryopreservation.
Sections du résumé
BACKGROUND
BACKGROUND
Peripheral blood stem cells (PBSC) are commonly cryopreserved awaiting clinical use for hematopoietic stem cell transplant. Long term cryopreservation is commonly defined as five years or longer, and limited data exists regarding how long PBSC can be cryopreserved and retain the ability to successfully engraft. Clinical programs, stem cell banks, and regulatory and accrediting agencies interested in product stability would benefit from such data. Thus, we assessed recovery and colony forming ability of PBSC following long-term cryopreservation as well as their ability to engraft in NOD/SCID/IL-2Rγ
AIM
OBJECTIVE
To investigate the in vivo engraftment potential of long-term cryopreserved PBSC units.
METHODS
METHODS
PBSC units which were collected and frozen using validated clinical protocols were obtained for research use from the Cellular Therapy Laboratory at Indiana University Health. These units were thawed in the Cellular Therapy Laboratory using clinical standards of practice, and the pre-freeze and post-thaw characteristics of the units were compared. Progenitor function was assessed using standard colony-forming assays. CD34-selected cells were transplanted into immunodeficient mice to assess stem cell function.
RESULTS
RESULTS
Ten PBSC units with mean of 17 years in cryopreservation (range 13.6-18.3 years) demonstrated a mean total cell recovery of 88% ± 12% (range 68%-110%) and post-thaw viability of 69% ± 17% (range 34%-86%). BFU-E growth was shown in 9 of 10 units and CFU-GM growth in 7 of 10 units post-thaw. Immunodeficient mice were transplanted with CD34-selected cells from four randomly chosen PBSC units. All mice demonstrated long-term engraftment at 12 wk with mean 34% ± 24% human CD45+ cells, and differentiation with presence of human CD19+, CD3+ and CD33+ cells. Harvested bone marrow from all mice demonstrated growth of erythroid and myeloid colonies.
CONCLUSION
CONCLUSIONS
We demonstrated engraftment of clinically-collected and thawed PBSC following cryopreservation up to 18 years in NSG mice, signifying likely successful clinical transplantation of PBSC following long-term cryopreservation.
Identifiants
pubmed: 32547684
doi: 10.4252/wjsc.v12.i5.359
pmc: PMC7280863
doi:
Types de publication
Journal Article
Langues
eng
Pagination
359-367Informations de copyright
©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict-of-interest statement: Goebel WS receives fees as a consulting medical director for Cook Regentec, LLC, and serves as medical director for Ossium Health, Inc. All other authors report no potential conflicts of interest.
Références
Bone Marrow Transplant. 2017 Dec;52(12):1599-1601
pubmed: 28650454
Biol Blood Marrow Transplant. 2015 Jan;21(1):50-4
pubmed: 25262882
Clin Cancer Res. 2011 Apr 15;17(8):2195-206
pubmed: 21487065
Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):645-50
pubmed: 12518050
Bone Marrow Res. 2011;2011:252953
pubmed: 22046557
Cryobiology. 2002 Jun;44(3):210-7
pubmed: 12237086
Adv Hematol. 2011;2011:517561
pubmed: 22190942
Blood. 2011 May 5;117(18):4773-7
pubmed: 21393480
Transfusion. 2010 Apr;50(4):808-19
pubmed: 19912586
Bone Marrow Transplant. 1996 Mar;17(3):425-30
pubmed: 8704699
Blood. 2010 Jul 15;116(2):193-200
pubmed: 20404133
Bone Marrow Transplant. 2014 Aug;49(8):1109-12
pubmed: 24797184
Stem Cells Transl Med. 2019 Apr;8(4):340-343
pubmed: 30843653
Bone Marrow Transplant. 2014 Jun;49(6):780-5
pubmed: 24686987
Cytotherapy. 2011 Aug;13(7):856-63
pubmed: 21385094
Bone Marrow Transplant. 1992 Jun;9(6):487-90
pubmed: 1628134
Transfusion. 2009 Aug;49(8):1709-19
pubmed: 19392777
Cytotherapy. 2012 Nov;14(10):1228-34
pubmed: 22900962
Cytotherapy. 2014 Jul;16(7):965-75
pubmed: 24910385
Oncol Rep. 2003 Nov-Dec;10(6):1993-8
pubmed: 14534732
Bone Marrow Transplant. 2007 Nov;40(9):831-5
pubmed: 17724443
Int J Hematol. 2011 Jan;93(1):99-105
pubmed: 21207212
Stem Cell Res Ther. 2015 Apr 17;6:64
pubmed: 25889496
Transplantation. 1981 Jun;31(6):454-7
pubmed: 7256827
Biol Blood Marrow Transplant. 2017 Apr;23(4):684-690
pubmed: 28013016
Clin Exp Immunol. 1997 Jan;107 Suppl 1:45-53
pubmed: 9020936
Biol Blood Marrow Transplant. 2010 Jan;16(1):102-7
pubmed: 19772946
Blood. 2012 Sep 27;120(13):2620-30
pubmed: 22517906
Bone Marrow Transplant. 2013 Jan;48(1):32-5
pubmed: 22659683