Debbarma Parishma, Mondal Tanmay, Manna Camelia, Kumar Kuldeep, Mukherjee Joydip, Das Bikash Chandra, Bag Sadhan, Das Kinsuk
Department of Veterinary Physiology, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.
Vet World. 2020 Dec;13(12):2772-2779. doi: 10.14202/vetworld.2020.2772-2779. Epub 2020 Dec 25.
Veterinary health care is an emergent area in animal sciences and innovative therapeutic approaches happen to be imperative in the present days. In view of the importance of cattle health and production, it is necessary to take up contemporary approach of stem cell therapy in this sector also. This study aimed to standardize an explant culture method of bovine umbilical tissue offcut to isolate mesenchymal stem cells (MSCs) because considerable efforts are required for ensuring easy accessibility and availability of MSCs in bulk quantity, as well as in establishing and characterizing the cell lines.
The umbilical cord (UC) tissue matrix offcut was collected after calving. A simplified cell isolation technique was followed to collect the emerged out cells from the explants of UC. Further, we expanded these isolated cells , observed its growth kinetics, and characterized to confirm as per the criterion of bovine MSCs.
A considerable exponential growth rate of the UC-derived cells was noticed. In addition to their confirmation as MSCs, the cells also exhibited plastic adherent property and maintained the spindle-shaped morphology throughout the culture. The cultured cells were found positive MSC-specific surface markers CD105, CD90, and CD73 and were negative for hematopoietic cell marker CD45. Cytochemical studies revealed the ability of the cells to differentiate into osteogenic, chondrogenic, and adipogenic lineages.
This simplified method of isolation and culture of bovine multipotent MSCs from the UC offcut collected after calving could be extrapolated for the greater availability of the cells for prospective therapeutic applications.
兽医保健是动物科学中的一个新兴领域,创新治疗方法在当今至关重要。鉴于牛健康和生产的重要性,在该领域采用当代干细胞治疗方法也很有必要。本研究旨在规范从牛脐带组织切下部分进行外植体培养的方法,以分离间充质干细胞(MSCs),因为要确保大量且容易获取MSCs,并建立和鉴定细胞系,需要付出相当大的努力。
产犊后收集脐带(UC)组织基质切下部分。采用一种简化的细胞分离技术从UC外植体中收集长出的细胞。此外,我们对这些分离出的细胞进行扩增,观察其生长动力学,并根据牛MSCs的标准进行鉴定。
观察到UC来源的细胞有相当高的指数生长率。除了确认为MSCs外,这些细胞还表现出贴壁特性,并且在整个培养过程中保持纺锤形形态。培养的细胞被发现MSCs特异性表面标志物CD105、CD90和CD73呈阳性,造血细胞标志物CD45呈阴性。细胞化学研究显示这些细胞具有分化为成骨、成软骨和成脂谱系的能力。
这种从产犊后收集的UC切下部分分离和培养牛多能MSCs的简化方法,可推广应用,以便为未来的治疗应用提供更多的细胞。