在无血清培养条件下扩增用于肝脏再生治疗的人类异体肝源祖细胞

IF 3.7 3区医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Cytotherapy Pub Date : 2024-07-14 DOI:10.1016/j.jcyt.2024.07.008

Pauline De Berdt , Elodie Deltour , Eric Pauly , Noelia Gordillo , Frédéric Lin , Etienne Sokal , Mustapha Najimi

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引用次数: 0

摘要

人异体肝源性祖细胞（HALPCs）具有向肝细胞样细胞分化的高级能力，并表现出强大的免疫调节、抗炎和抗纤维化特性。HALPCs 已按照良好生产规范 (GMP) 成功生产，目前正在进行临床开发。之前的一项 2a 期试验证明了外周静脉注射 HALPCs 的安全性，并初步证明了这种细胞具有恢复急性-慢性肝功能衰竭 (ACLF) 患者肝功能的特性，从而有可能改善他们的生存状况。目前正在多个中心进行 2b 期试验（NCT04229901），以获得疗效和安全性方面的概念验证。HALPCs 目前使用胎牛血清 (FBS) 生产，这可能会导致不同批次产品的质量和数量存在差异。使用无血清培养基（SFM）是克服这种变异性的另一种方法，同时也完全符合法规要求。本研究的目的是将目前含 FBS 的培养条件与两种符合 GMP 标准的工业 SFM 进行比较：StemMACS（Miltenyi Biotec，Bergisch Gladbach，德国）和 PRIME-XV（FUJIFILM Irvine Scientific，Santa Ana，美国加利福尼亚州）都能显著刺激 HALPCs 的增殖，缩短其萌发期和种群倍增时间。通过常规核型分析，这一效果与 HALPCs 遗传稳定性的显著提高相关。流式细胞术和酶联免疫吸附试验（ELISA）分别证明，在 SFM 中培养的 HALPCs 的表达谱（特性和纯度）和功能均得以保持。通过前列腺素 E2（PGE2）分泌对其有效性进行评估，结果显示，在 FBS 和 SFM 条件下，HALPCs 对 CD4+ T 细胞增殖的影响相似。此外，与 FBS 条件相比，SFM 条件下培养的 HALPCs 中更大比例的组织因子（CD142）表达增强。

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Expansion of human allogeneic liver-derived progenitor cells for liver regenerative therapy in serum-free culture conditions

Human allogeneic liver-derived progenitor cells (HALPCs) display advanced ability to differentiate into hepatocyte-like cells and exhibit potent immunomodulatory, anti-inflammatory, and anti-fibrotic properties. HALPCs have been successfully manufactured under good manufacturing practice (GMP) and are currently in clinical development. A previous phase 2a trial demonstrated the safety of peripheral intravenous infusions of HALPCs and preliminary evidence of the cells’ properties to restore liver function in patients with acute-on-chronic liver failure (ACLF), thus potentially improving their survival. A phase 2b trial is currently ongoing across multiple centers (NCT04229901) to obtain proof-of-concept on efficacy and additional safety. HALPCs are currently manufactured using fetal bovine serum (FBS), which can reveal qualitative and quantitative variations between batches. The use of serum-free medium (SFM) represents an alternative means to overcome this variability while also complying fully with regulations. The aim of this study was to compare current FBS-containing culture conditions with two industry-available GMP-compliant SFMs: StemMACS (Miltenyi Biotec, Bergisch Gladbach, Germany) and PRIME-XV (FUJIFILM Irvine Scientific, Santa Ana, California, USA).

The proliferation of HALPCs was significantly stimulated by both SFMs, which shortened both their emergence period and population doubling time. This effect was correlated with a significant improvement in their genetic stability as analyzed by conventional karyotyping. The expression profile (identity and purity) and functionality of HALPCs cultured in SFM were maintained, as demonstrated by flow cytometry and enzyme-linked immunoassay (ELISA), respectively. Their potency, evaluated via prostaglandin E2 (PGE2) secretion, showed a similar effect on CD4⁺ T-cell proliferation in FBS and SFM conditions. Furthermore, a greater proportion of HALPCs cultured in SFM showed enhanced expression of tissue factor (CD142) compared with the FBS condition.

Altogether, SFM conditions enabled consistent HALPC quality to be achieved without altering their expression and functional profiles.

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来源期刊

Cytotherapy 医学-生物工程与应用微生物

CiteScore

6.30

自引率

4.40%

发文量

683

审稿时长

49 days

期刊介绍： The journal brings readers the latest developments in the fast moving field of cellular therapy in man. This includes cell therapy for cancer, immune disorders, inherited diseases, tissue repair and regenerative medicine. The journal covers the science, translational development and treatment with variety of cell types including hematopoietic stem cells, immune cells (dendritic cells, NK, cells, T cells, antigen presenting cells) mesenchymal stromal cells, adipose cells, nerve, muscle, vascular and endothelial cells, and induced pluripotential stem cells. We also welcome manuscripts on subcellular derivatives such as exosomes. A specific focus is on translational research that brings cell therapy to the clinic. Cytotherapy publishes original papers, reviews, position papers editorials, commentaries and letters to the editor. We welcome "Protocols in Cytotherapy" bringing standard operating procedure for production specific cell types for clinical use within the reach of the readership.