PMID- 36619595
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230111
IS  - 2306-9759 (Print)
IS  - 2313-0792 (Electronic)
IS  - 2306-9759 (Linking)
VI  - 9
DP  - 2022
TI  - Creation of human hematopoietic chimeric cell (HHCC) line as a novel strategy for 
      tolerance induction in transplantation.
PG  - 11
LID - 10.21037/sci-2022-026 [doi]
LID - 11
AB  - BACKGROUND: Cell-based and chimerism-based therapies represent a promising 
      approach for tolerance induction in transplantation. We propose a new cell 
      therapy of the ex vivo created human hematopoietic chimeric cells (HHCC) as an 
      alternative approach to bone marrow (BM)-based therapies in support of solid 
      organ and vascularized composite allotransplantation (VCA). This study aimed to 
      characterize in vitro the phenotype, genotype, clonogenic, and tolerogenic 
      properties of HHCC. METHODS: Thirty ex vivo fusions of CD34(+) cells from two 
      unrelated human BM donors were performed. CD34(+) cells were stained separately 
      with PKH26 and PKH67 membrane dyes and fused using polyethylene glycol (PEG). 
      Creation of human HHCC and chimeric state was confirmed by flow cytometry (FC), 
      confocal microscopy (CM) and electron microscopy (EM). HHCC's phenotype (CD34, 
      CD133, CD117, CD4, CD19, CD4/CD25) was assessed by FC, viability by Trypan Blue, 
      LIVE/DEAD and apoptosis by AnnexinV/Sytox Blue and TUNEL assay, while mixed 
      lymphocyte reaction (MLR) assay assessed HHCC's immunogenicity and tolerogenic 
      properties. HHCC differentiation, proliferation and clonogenic potential were 
      assessed by the colony forming unit (CFU). Polyploidy was evaluated by 
      fluorescence in situ hybridization (FISH), whereas polymerase chain 
      reaction-reverse sequence-specific oligonucleotide probe (PCR-rSSOP) and short 
      tandem repeats-polymerase chain reaction (STR-PCR) assessed HHCC's genotype, and 
      chimerism. Reverse transcription polymerase chain reaction (RT-PCR) analyzed 
      cytokines secretion [interleukin (IL)-10, transforming growth factor-beta (TGF-beta) 
      and tumor necrosis factor-alpha (TNF-alpha)] up to 14 days post-fusion. RESULTS: FC and 
      CM confirmed creation of HHCC by fusion of CD34(+) cells from two unrelated human 
      donors. After fusion, maintenance of hematopoietic markers and increased 
      expression of Treg-cells (CD4/CD25) was confirmed. Moreover, high HHCC viability 
      (99%) and a low apoptosis rate (1.2%) were revealed HHCC presented decreased 
      immunogenicity by MLR, and significant, 40-fold increase of IL-10 the 
      pro-tolerogenic cytokine at 21 days after fusion (RT-PCR) P<0.0001. The number of 
      polyploid cells was negligible (0.48%). PCR-rSSOP of HHCC after fusion confirmed 
      presence of human leukocyte antigen (HLA) class I and class II-alleles and 
      presence of the loci specific for both CD34(+) cells BM donors by STR-PCR. 
      CONCLUSIONS: We have created a new hematopoietic cell line of HHCC from two 
      unrelated human donors, and have successfully characterized in vitro, viability, 
      phenotype, genotype, clonogenic, and tolerogenic properties of HHCC. These unique 
      immunomodulatory and tolerogenic properties introduce HHCC as a novel therapeutic 
      approach for tolerance induction in VCA and solid organ transplantation.
CI  - 2022 Stem Cell Investigation. All rights reserved.
FAU - Siemionow, Maria
AU  - Siemionow M
AD  - Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL, USA.
FAU - Brodowska, Sonia
AU  - Brodowska S
AD  - Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL, USA.
FAU - Rozczka, Klaudia
AU  - Rozczka K
AD  - Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL, USA.
FAU - Roesler, Claire
AU  - Roesler C
AD  - Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL, USA.
LA  - eng
PT  - Journal Article
DEP - 20221219
PL  - China
TA  - Stem Cell Investig
JT  - Stem cell investigation
JID - 101672113
PMC - PMC9813662
OTO - NOTNLM
OT  - Human hematopoietic chimeric cells therapy (HHCC therapy)
OT  - allograft rejection
OT  - chimerism
OT  - tolerance induction
OT  - vascularized composite allotransplantation (VCA)
COIS- Conflicts of Interest: All authors have completed the ICMJE uniform disclosure 
      form (available at 
      https://sci.amegroups.com/article/view/10.21037/sci-2022-026/coif). MS reports 
      consulting fees from AMCA. The other authors have no conflicts of interest to 
      declare.
EDAT- 2023/01/10 06:00
MHDA- 2023/01/10 06:01
PMCR- 2022/12/19
CRDT- 2023/01/09 03:54
PHST- 2022/08/23 00:00 [received]
PHST- 2022/11/21 00:00 [accepted]
PHST- 2023/01/09 03:54 [entrez]
PHST- 2023/01/10 06:00 [pubmed]
PHST- 2023/01/10 06:01 [medline]
PHST- 2022/12/19 00:00 [pmc-release]
AID - sci-09-2022-026 [pii]
AID - 10.21037/sci-2022-026 [doi]
PST - epublish
SO  - Stem Cell Investig. 2022 Dec 19;9:11. doi: 10.21037/sci-2022-026. eCollection 
      2022.