Establishment and characterization of hTERT-immortalized porcine muscle stem cells, and their prospective uses

IF 3.1 3区农林科学 Q3 FOOD SCIENCE & TECHNOLOGY Food Science and Biotechnology Pub Date : 2024-12-11 DOI:10.1007/s10068-024-01785-9

Witsanu Srila, Amorn Pangjantuk, Phongsakorn Kunhorm, Nipha Chaicharoenaudomrung, Parinya Noisa

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Abstract

Myogenic differentiation ability made porcine muscle satellite cells (MSCs) promising candidates for cultured meat production. While in vitro, porcine primary cells lose differentiation capacity, have short lifespans, and change phenotype. For immortal porcine MSCs, human telomerase reverse transcriptase (hTERT) gene was overexpressed in parental cells to restore telomerase activity and lengthen cell longevity. After selection, G418-resistant cells were expanded and passed by different generations. The hTERT-immortalized MSCs presented spindle-like shape, telomere extension, and indefinite proliferation. In comparison to parental cells, immortal cells grew more rapidly and doubled faster. Immortal MSCs expressed muscle-specific protein and gene markers, were self-renewing stem cells, and could develop into myofibers in vitro. In culture plates with more than 100 generations, immortal MSCs formed tumors, but not lower passaged cells. Today, we showed that hTERT can immortalize primary porcine MSCs and preserve their stem cell characteristics. For research and cultured meat technologies, immortality may be valuable.

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htert永生化猪肌肉干细胞的建立、表征及其应用前景

猪肌肉卫星细胞（间充质干细胞）的肌源分化能力使其有望成为肉类生产的候选培养细胞。在体外培养时，猪原代细胞会丧失分化能力、寿命短并改变表型。为了获得永生的猪间叶干细胞，我们在亲代细胞中过表达人类端粒酶逆转录酶（hTERT）基因，以恢复端粒酶活性并延长细胞寿命。经过选择后，抗G418的细胞被扩增并通过不同的世代传承。经 hTERT 处死的间充质干细胞呈纺锤形，端粒延长，无限增殖。与亲代细胞相比，永生细胞生长更快，翻倍更快。永生间充质干细胞表达肌肉特异性蛋白和基因标记，是自我更新的干细胞，可在体外发育成肌纤维。在超过100代的培养板中，永生间充质干细胞会形成肿瘤，而低传代细胞则不会。今天，我们证明了 hTERT 能使猪原代间充质干细胞永生，并保留其干细胞特性。对于研究和培养肉类技术来说，永生可能很有价值。

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来源期刊

Food Science and Biotechnology FOOD SCIENCE & TECHNOLOGY-

CiteScore

5.40

自引率

3.40%

发文量

174

审稿时长

2.3 months

期刊介绍： The FSB journal covers food chemistry and analysis for compositional and physiological activity changes, food hygiene and toxicology, food microbiology and biotechnology, and food engineering involved in during and after food processing through physical, chemical, and biological ways. Consumer perception and sensory evaluation on processed foods are accepted only when they are relevant to the laboratory research work. As a general rule, manuscripts dealing with analysis and efficacy of extracts from natural resources prior to the processing or without any related food processing may not be considered within the scope of the journal. The FSB journal does not deal with only local interest and a lack of significant scientific merit. The main scope of our journal is seeking for human health and wellness through constructive works and new findings in food science and biotechnology field.