中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (31): 8092-8099.doi: 10.12307/2026.450

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

间充质干细胞条件培养基精华液的制备及性能评价

王泉生1，2，3，陈松涛1，2，3，张永清1，2，3，乔  华1，2，3   

  1. 1山西医科大学基础医学院生物化学与分子生物学教研室，山西省太原市   030001；2煤炭环境致病与防治教育部重点实验室，山西省太原市   030001；3出生缺陷与细胞再生山西省重点实验室，山西省太原市   030001
  • 收稿日期:2025-12-01 接受日期:2026-02-02 出版日期:2026-11-08 发布日期:2026-05-22
  • 通讯作者: 乔华，博士，副教授，硕士生导师，山西医科大学基础医学院生物化学与分子生物学教研室，山西省太原市 030001；煤炭环境致病与防治教育部重点实验室，山西省太原市 030001；出生缺陷与细胞再生山西省重点实验室，山西省太原市 030001
  • 作者简介:王泉生，男，2002年生，山西医科大学医学科学院在读硕士，主要从事干细胞衍生物治疗方面的研究。
  • 基金资助:
    山西省大学生创新创业计划项目(20240411)，指导教师：乔华；山西省自然科学基金项目(202203021211232)，项目负责人：乔华

Preparation and performance evaluation of essence with mesenchymal stem cell conditioned medium

Wang Quansheng1, 2, 3, Chen Songtao1, 2, 3, Zhang Yongqing1, 2, 3, Qiao Hua1, 2, 3   

  1. 1Department of Biochemistry and Molecular Biology, School of Basic Medicine, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 2Ministry of Education Key Laboratory of Coal Environmental Pathogenicity and Prevention, Taiyuan 030001, Shanxi Province, China; 3Shanxi Key Laboratory of Birth Defect and Cell Regeneration, Taiyuan 030001, Shanxi Province, China
  • Received:2025-12-01 Accepted:2026-02-02 Online:2026-11-08 Published:2026-05-22
  • Contact: Qiao Hua, MD, Associate professor, Master’s supervisor, Department of Biochemistry and Molecular Biology, School of Basic Medicine, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; Ministry of Education Key Laboratory of Coal Environmental Pathogenicity and Prevention, Taiyuan 030001, Shanxi Province, China; Shanxi Key Laboratory of Birth Defect and Cell Regeneration, Taiyuan 030001, Shanxi Province, China
  • About author:Wang Quansheng, MS candidate, Department of Biochemistry and Molecular Biology, School of Basic Medicine, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; Ministry of Education Key Laboratory of Coal Environmental Pathogenicity and Prevention, Taiyuan 030001, Shanxi Province, China; Shanxi Key Laboratory of Birth Defect and Cell Regeneration, Taiyuan 030001, Shanxi Province, China
  • Supported by:
    Shanxi Provincial Undergraduate Innovation and Entrepreneurship Program, No. 20240411 (to QH); Shanxi Provincial Natural Science Foundation Project, No. 202203021211232 (to QH)

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摘要：

文题释义：

间充质干细胞：是一类存在于人体多种组织(如骨髓、脂肪、脐带等)中的“多能”成体干细胞，具有多向分化潜能，如分化为成骨细胞、脂肪细胞等，同时间充质干细胞分泌大量生物活性物质，包括生长因子、细胞因子、外泌体等，这些物质能抑制细胞凋亡、刺激内源性祖细胞增殖分化，并具有抗纤维化、抗炎等作用，在再生医学和免疫调控领域有广阔的临床应用前景。
条件培养基：是培养细胞后收集的培养基，其中富含细胞分泌的全套生物活性物质，即“分泌组”，包括生长因子、细胞因子、趋化因子、代谢产物以及作为细胞间通讯关键载体的外泌体等。基于条件培养基能模拟复杂的体内微环境，在肿瘤微环境探索、组织再生等领域得到了广泛应用。

摘要
背景：间充质干细胞条件培养基含有多种生物活性物质，具有减缓人皮肤成纤维细胞衰老的潜力，但关于间充质干细胞条件培养基的应用研究较少。
目的：制备人脐带间充质干细胞条件培养基精华液并进行性能评价。
方法：通过感官性能评分设计正交试验筛选间充质干细胞条件培养基精华液配方，通过4 ℃条件贮存稳定性实验、冷热循环稳定性实验、离心稳定性实验评估精华液的稳定性，从溶血实验、细胞毒性实验、皮肤刺激实验3个层面评价精华液的使用安全性，采用保湿实验检测失水率、CCK-8法检测细胞活性，以及利用人皮肤成纤维细胞及斑马鱼胚胎衰老模型检测活性氧水平考察精华液的保湿和抗衰老功效。
结果与结论：①正交试验筛选得到间充质干细胞条件培养基添加量为5%、甘油添加量为5%、黄原胶添加量为0.25%时，感官性能评分最高，且配方重复性好，可行性高；②间充质干细胞条件培养基精华液在4 ℃贮存、冷热循环及离心条件下的稳定性良好、保湿性能佳；③间充质干细胞条件培养基精华液的血液相容性好、细胞毒性低，无皮肤刺激作用，安全可靠；④间充质干细胞条件培养基精华液能促进衰老人皮肤成纤维细胞增殖，降低人皮肤成纤维细胞及斑马鱼胚胎衰老模型的活性氧水平，表现出一定的抗衰老、抗氧化和保湿性能。以上表明，以间充质干细胞条件培养基为成分制备的精华液稳定性好、温和无刺激且具有保湿、抗衰老和抗氧化功效。

关键词: 间充质干细胞, 条件培养基, 复制性衰老, 抗衰老, 保湿, 抗氧化, 精华液, 人脐带间充质干细胞, 干细胞衍生物

Abstract: BACKGROUND: Mesenchymal stem cell conditioned medium contains a variety of bioactive substances, and has the potential to slow down the aging of skin fibroblasts. However, there are few studies on the practical application of mesenchymal stem cell conditioned medium.
OBJECTIVE: To prepare and evaluate the performance of an essence containing human umbilical cord mesenchymal stem cell-conditioned medium. 
METHODS: The formulas of mesenchymal stem cell conditioned medium essence were screened by orthogonal experiment design using sensory performance score. The stability of the essence was evaluated through storage stability test, thermal cycling stability test, and centrifugal stability test at 4°C. The safety of the essence was evaluated from three levels: hemolysis test, cytotoxicity test, and skin irritation test. The moisturizing and anti-aging effects of the serum were investigated by measuring water loss rate using moisturizing test, cell viability using the CCK-8 assay, and reactive oxygen species levels using human skin fibroblasts and a zebrafish embryo aging model. 
RESULTS AND CONCLUSION: (1) Orthogonal experimental design showed that when the addition of mesenchymal stem cell conditioned medium was 5%, glycerol was 5%, and xanthan gum was 0.25%, the sensory performance score was the highest, and the formula had good repeatability and high feasibility. (2) The mesenchymal stem cell conditioned medium essence had good stability under the conditions of 4 °C storage, hot and cold cycling and centrifugation condition, and low water loss rate. (3) The mesenchymal stem cell conditioned medium essence had good hemocompatibility, low cytotoxicity, and no skin irritation, and was safe and reliable. (4) The mesenchymal stem cell conditioned medium essence could promote the proliferation of senescent skin fibroblasts cells and reduce the levels of reactive oxygen species in aging model cells and zebrafish embryos, indicating certain anti-aging, antioxidant effects, and moisturizing properties. The above results show that the essence prepared with mesenchymal stem cell conditioned medium as the functional ingredient has good stability, is mild and non-irritating, and has anti-aging, moisturizing, and antioxidant effects.

Key words: mesenchymal stem cells, conditioned medium, replicative senescence, anti-aging, moisturizing, antioxidant, essence, human umbilical cord mesenchymal stem cells, stem cell derivatives

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