Safety Assessment of Autologous Stem Cell Combination Therapy in Patients With Decompensated Liver Cirrhosis: A Pilot Study

Abstract

Background: Haematopoietic stem cell (HSC) infusion has demonstrated short-term improvement in liver functions in patients with chronic liver disease. The combination of HSC with mesenchymal stem cells (MSCs), which has an immunomodulatory effect, may augment the effects and enhance the duration of improvements on liver functions. The aim of the present study was to assess the safety of infusing the combination of autologous HSCs and MSCs in decompensated liver cirrhosis.

Methods: In phase I of the study, in vitro assessment was performed to observe the effect of coculturing MSCs with HSCs on their viability and cytokine profiles. Phase II of the study was to assess the safety of combination of stem cell infusions. Bone marrow (50 ml) was aspirated for MSC isolation and expansion using standard protocol. Patients received subcutaneous doses (n = 5) of granulocyte colony-stimulating factor (G-CSF) for stem cell mobilization followed by leukapheresis for harvesting HSCs using CliniMacs. HSCs and MSCs were infused through the hepatic artery under fluoroscopic guidance and were monitored for any adverse effects.

Results: In vitro studies revealed 94% viable HSCs in coculture similar to monoculture. HSCs released only interleukin (IL)-8, whereas MSCs secreted IL-8 and IL-6 in monocultures, and both IL-8 and IL-6 were secreted in coculture. G-CSF administration- and bone marrow aspiration-related complications were not observed. Infusion of the cells through the hepatic artery was safe, and no postprocedural complications were noted.

Conclusion: The combination of autologous HSC and MSC infusion is a safe procedure in patients with decompensated liver cirrhosis, and the outcomes needed to be assessed in larger studies.

Trial number: NCT04243681.

Keywords: 7-AAD, 7-aminoactinomycin D; AFP, alpha-fetoprotein; CBA, cytokine cytometric bead assay; CLD, chronic liver disease; DMEM-KO, Dulbecco's modified Eagle's Knock out medium; FBS, foetal bovine serum; G-CSF, granulocyte colony-stimulating factor; HSC, haematopoietic stem cell; IL, interleukin; MELD, Model for End-Stage Liver Disease; MNC, mononuclear cell; MSC, mesenchymal stem cell; SOP, standard operating procedure; TJLB, transjugular liver biopsy; USG, ultrasonography; cath-lab, cardiac catheterization laboratory; cirrhosis of liver; combination of stem cells; mesenchymal stem cells; stem cells.

© 2021 Indian National Association for Study of the Liver. Published by Elsevier B.V. All rights reserved.

Figures

Figure 1

Representative flow cytometry pictures of the healthy control (upper panel) and patient with CLD (lower panel). Panel a shows gating of MSCs at passage 3. Panel b shows MSCs positive for phenotypic marker staining for CD90, CD73 and CD105 and negative for CD34, CD45 and viability marker 7-AAD. 7-AAD, 7-aminoactinomycin D; MSC, mesenchymal stem cell; CLD, chronic liver disease.

Figure 2

Trilineage differentiation of BM-derived MSCs in healthy controls and patients with CLD. BM-derived MSCs characterized for differentiation into adipocytes (formation of lipid droplets stained with Oil Red), osteocytes (demonstrated by calcium deposition stained with Alizarin Red) and chondrocytes (demonstrated by the deposition of extracellular matrix stained with toluidine blue) from healthy controls (A) and (B) patients with CLD. CLD, chronic liver disease; BM, bone marrow; MSC, mesenchymal stem cell.

Figure 3

Cocultures of MSCs and CD34+ stem cells. Monoculture of MSCs (A) and HSCs (B) and coculture of MSCs and HSCs (C) at 72 h demonstrate 99% viable HSCs in cocultures. HSC, haematopoietic stem cell; MSC, mesenchymal stem cell.

Figure 4

Cytokine analysis of culture supernatants of BM-derived stem cells from patients with CLD. Cytokine analysis from the BM-derived MSC culture supernatant shows IL-8 and IL-6 at 24 h and 72 h. The CD34+ culture supernatant secretes only IL-8. The coculture supernatants secrete IL-8 and IL-6. IL, interleukin; CLD, chronic liver disease; MSC, mesenchymal stem cell; BM, bone marrow.