Chijimatsu Ryota, Takeda Taiga, Tsuji Shinsaku, Sasaki Kohei, Kato Koichi, Kojima Rie, Michihata Noriko, Tsubaki Toshiya, Matui Aya, Watanabe Miharu, Tanaka Sakae, Saito Taku
The University of Tokyo, Bone and Cartilage Regenerative Medicine, Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
The University of Tokyo, Sensory and Motor System Medicine, Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan.
Regen Ther. 2022 Jun 8;21:52-61. doi: 10.1016/j.reth.2022.05.009. eCollection 2022 Dec.
Adipose-derived stem cells (ASCs) are an attractive cell source for cell therapy. Despite the increasing number of clinical applications, the methodology for ASC isolation is not optimized for every individual. In this study, we developed an effective material to stabilize explant cultures from small-fragment adipose tissues.
Polypropylene/polyethylene nonwoven sheets were coated with hydroxyapatite (HA) particles. Adipose fragments were then placed on these sheets, and their ability to trap tissue was monitored during explant culture. The yield and properties of the cells were compared to those of cells isolated by conventional collagenase digestion.
Hydroxyapatite-coated nonwovens immediately trapped adipose fragments when placed on the sheets. The adhesion was stable even in culture media, leading to cell migration and proliferation from the tissue along with the nonwoven fibers. A higher fiber density further enhanced cell growth. Although cells on nonwoven explants could not be fully collected with cell dissociation enzymes, the cell yield was significantly higher than that of conventional monolayer culture without impacting stem cell properties.
Hydroxyapatite-coated nonwovens are useful for the effective primary explant culture of connective tissues without enzymatic cell dissociation.
脂肪来源干细胞(ASCs)是细胞治疗中一种有吸引力的细胞来源。尽管临床应用数量不断增加,但ASC分离方法并非针对每个个体进行了优化。在本研究中,我们开发了一种有效的材料来稳定小片段脂肪组织的外植体培养。
聚丙烯/聚乙烯无纺布片材涂覆有羟基磷灰石(HA)颗粒。然后将脂肪片段放置在这些片材上,并在体外培养期间监测其捕获组织的能力。将细胞的产量和特性与通过传统胶原酶消化分离的细胞进行比较。
涂覆有羟基磷灰石的无纺布在放置在片材上时立即捕获脂肪片段。即使在培养基中,粘附也很稳定,导致细胞从组织沿着无纺布纤维迁移和增殖。更高的纤维密度进一步促进了细胞生长。尽管无纺布外植体上的细胞不能用细胞解离酶完全收集,但细胞产量明显高于传统单层培养,且不影响干细胞特性。
涂覆有羟基磷灰石的无纺布可用于结缔组织的有效原代外植体培养,无需酶促细胞解离。