PMID- 37614644
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230825
IS  - 2306-9759 (Print)
IS  - 2313-0792 (Electronic)
IS  - 2306-9759 (Linking)
VI  - 10
DP  - 2023
TI  - Novel Human Umbilical Di-Chimeric (HUDC) cell therapy for transplantation without 
      life-long immunosuppression.
PG  - 16
LID - 10.21037/sci-2023-024 [doi]
LID - 16
AB  - BACKGROUND: Cell-based therapies are promising for tolerance induction in bone 
      marrow (BM), solid organs, and vascularized composite allotransplantation (VCA). 
      The toxicity of bone marrow transplantation (BMT) protocols precludes this 
      approach from routine clinical applications. To address this problem, we 
      developed a new therapy of Human Umbilical Di-Chimeric (HUDC) cells for tolerance 
      induction in transplantation. This study established in vitro characterization of 
      the created HUDC cells. METHODS: We performed sixteen ex vivo polyethylene glycol 
      (PEG)-mediated fusions of human umbilical cord blood (UCB) cells from two 
      unrelated donors. Fusion feasibility was confirmed in vitro by flow cytometry 
      (FC) and confocal microscopy (CM). The HUDC cells' genotype was assessed by 
      lymphocytotoxicity test and short tandem repeat-polymerase chain reaction 
      (STR-PCR) analysis, phenotype by FC, viability by LIVE/DEAD((R)) assay, and 
      apoptosis level by Annexin V staining. We used COMET assay to assess HUDC cells' 
      genotoxicity after the fusion procedure. Clonogenic properties of HUDC cells were 
      evaluated by colony forming unit (CFU) assay. Mixed lymphocyte reaction (MLR) 
      assay assessed immunogenic and tolerogenic properties of HUDC cells. RESULTS: We 
      confirmed the creation of HUDC cells from two unrelated human donors of UCB cells 
      by FC and CM. Human leukocyte antigen (HLA) class I and II typing, and STR-PCR 
      analysis of HUDC cells confirmed the presence of alleles and loci from both 
      unrelated UCB donors (donor chimerism: 49%+/-8.3%, n=4). FC confirmed the 
      hematopoietic phenotype of HUDC cells. We confirmed high HUDC cells' viability 
      (0.47% of dead cells) and a low apoptosis level of fused HUDC cells (15.9%) 
      compared to positive control of PKH-stained UCB cells (20.4%) before fusion. 
      COMET assay of HUDC cells revealed a lack of DNA damage. CFU assay confirmed 
      clonogenic properties of HUDC cells, and MLR assay revealed a low immunogenicity 
      of HUDC cells. CONCLUSIONS: This study confirmed creation of a novel HUDC cell 
      line by ex vivo PEG-mediated fusion of UCB cells from two unrelated donors. The 
      unique concept of creating a HUDC cell line, representing the genotype and 
      phenotype of both, transplant donor and the recipient, introduces a promising 
      approach for tolerance induction in BM, solid organs, and VCA transplantation.
CI  - 2023 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 - Cwykiel, Joanna
AU  - Cwykiel J
AD  - Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL, USA.
FAU - Chambily, Lucile
AU  - Chambily L
AD  - Department of Orthopaedics, University of Illinois at Chicago, Chicago, IL, USA.
FAU - Gacek, Stephanie
AU  - Gacek S
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.
LA  - eng
PT  - Journal Article
DEP - 20230814
PL  - China
TA  - Stem Cell Investig
JT  - Stem cell investigation
JID - 101672113
PMC - PMC10442563
OTO - NOTNLM
OT  - Chimerism
OT  - Human Umbilical Di-Chimeric cell therapy (HUDC cell therapy)
OT  - stem cells
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-2023-024/coif). The authors 
      have no conflicts of interest to declare.
EDAT- 2023/08/24 06:42
MHDA- 2023/08/24 06:43
PMCR- 2023/08/14
CRDT- 2023/08/24 04:05
PHST- 2023/04/30 00:00 [received]
PHST- 2023/08/02 00:00 [accepted]
PHST- 2023/08/24 06:43 [medline]
PHST- 2023/08/24 06:42 [pubmed]
PHST- 2023/08/24 04:05 [entrez]
PHST- 2023/08/14 00:00 [pmc-release]
AID - sci-10-2023-024 [pii]
AID - 10.21037/sci-2023-024 [doi]
PST - epublish
SO  - Stem Cell Investig. 2023 Aug 14;10:16. doi: 10.21037/sci-2023-024. eCollection 
      2023.