临床级软骨诱导性老年成体干细胞的货架寿命评估用于软骨再生。

Shelf Life Evaluation of Clinical Grade Chondrogenic Induced Aged Adult Stem Cells for Cartilage Regeneration.

机构信息

Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, K. L, Malaysia.

Bioartificial Organ and Regenerative Medicine Unit, National Defence University of Malaysia, Sungai Besi, Camp, 57000, K. L, Malaysia.

出版信息

Sci Rep. 2018 Mar 12;8(1):4345. doi: 10.1038/s41598-018-22748-1.

The study objectives include, enhancing the proliferations of aged bone marrow stem cells (BMSCs) and adipose stem cells (ADSCs); and evaluating the shelf lives of clinical grade chondrogenically induced cells from both samples. ADSCs and BMSCs from 56 patients (76 ± 8 yrs) were proliferated using basal medium (FD) and at (5, 10, 15, 20 and 25) ng/ml of basal fibroblast growth factor (bFGF). They were induced to chondrogenic lineage and stored for more than 120 hrs in FD, serum, Dulbecco's phosphate buffered saline (DPBS) and saline at 4 °C. In FD, cells stagnated and BMSCs' population doubling time (PDT) was 137 ± 30 hrs, while ADSCs' was 129.7 ± 40 hrs. bFGF caused PDT's decrease to 24.5 ± 5.8 hrs in BMSCs and 22.0 ± 6.5 hrs in ADSCs (p = 0.0001). Both cells were positive to stem cell markers before inductions and thereafter, expressed significantly high chondrogenic genes (p = 0.0001). On shelf life, both cells maintained viabilities and counts above 70% in FD and serum after 120 hrs. BMSCs' viabilities in DPBS fell below 70% after 96 hrs and saline after 72 hrs. ADSCs' viability fell below 70% in DPBS after 24 hrs and saline within 24 hrs. Concentrations between 20 ng/ml bFGF is ideal for aged adult cells' proliferation and delivery time of induced BMSCs and ADSCs can be 120 hrs in 4 °C serum.

研究目的包括，促进老年骨髓间充质干细胞（BMSCs）和脂肪间充质干细胞（ADSCs）的增殖；评估来自两种样本的临床级软骨诱导细胞的保质期。使用基础培养基（FD）和 5、10、15、20 和 25ng/ml 的基础成纤维细胞生长因子（bFGF）对来自 56 名患者（76±8 岁）的 ADSCs 和 BMSCs 进行增殖。将它们诱导为软骨谱系，并在 FD、血清、杜氏磷酸盐缓冲盐水（DPBS）和 4°C 的盐水中储存超过 120 小时。在 FD 中，细胞停滞不前，BMSCs 的倍增时间（PDT）为 137±30 小时，而 ADSCs 的 PDT 为 129.7±40 小时。bFGF 使 BMSCs 的 PDT 减少至 24.5±5.8 小时，ADSCs 的 PDT 减少至 22.0±6.5 小时（p=0.0001）。在诱导之前和之后，两种细胞均对干细胞标志物呈阳性，此后，它们都表达了显著高的软骨基因（p=0.0001）。在保质期方面，在 120 小时后，两种细胞在 FD 和血清中均保持活力和计数均高于 70%。BMSCs 在 DPBS 中的活力在 96 小时后低于 70%，在盐水中低于 70%在 72 小时后。ADSCs 在 DPBS 中的活力在 24 小时后低于 70%，在盐水中在 24 小时内低于 70%。20ng/ml 的 bFGF 浓度最适合老年成体细胞的增殖，诱导的 BMSCs 和 ADSCs 的输送时间可在 4°C 血清中保持 120 小时。