Liu Fei, Xiao Jing, Chen Lei-Hui, Pan Yu-Yue, Tian Jun-Zhang, Zhang Zhi-Ren, Bai Xiao-Chun
School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong Province, China.
Department of Health Management, Guangdong Second Provincial General Hospital, Guangzhou 510317, Guangdong Province, China.
World J Stem Cells. 2024 Mar 26;16(3):287-304. doi: 10.4252/wjsc.v16.i3.287.
The self-assembly of solid organs from stem cells has the potential to greatly expand the applicability of regenerative medicine. Stem cells can self-organise into microsized organ units, partially modelling tissue function and regeneration. Dental pulp organoids have been used to recapitulate the processes of tooth development and related diseases. However, the lack of vasculature limits the utility of dental pulp organoids.
To improve survival and aid in recovery after stem cell transplantation, we demonstrated the three-dimensional (3D) self-assembly of adult stem cell-human dental pulp stem cells (hDPSCs) and endothelial cells (ECs) into a novel type of spheroid-shaped dental pulp organoid under hypoxia and conditioned medium (CM).
During culture, primary hDPSCs were induced to differentiate into ECs by exposing them to a hypoxic environment and CM. The hypoxic pretreated hDPSCs were then mixed with ECs at specific ratios and conditioned in a 3D environment to produce prevascularized dental pulp organoids. The biological characteristics of the organoids were analysed, and the regulatory pathways associated with angiogenesis were studied.
The combination of these two agents resulted in prevascularized human dental pulp organoids (Vorganoids) that more closely resembled dental pulp tissue in terms of morphology and function. Single-cell RNA sequencing of dental pulp tissue and RNA sequencing of Vorganoids were integrated to analyse key regulatory pathways associated with angiogenesis. The biomarkers forkhead box protein O1 and fibroblast growth factor 2 were identified to be involved in the regulation of Vorganoids.
In this innovative study, we effectively established an model of Vorganoids and used it to elucidate new mechanisms of angiogenesis during regeneration, facilitating the development of clinical treatment strategies.
利用干细胞自组装形成实体器官，有望极大地拓展再生医学的应用范围。干细胞能够自我组织形成微型器官单元，部分模拟组织功能和再生过程。牙髓类器官已被用于重现牙齿发育及相关疾病的过程。然而，缺乏脉管系统限制了牙髓类器官的实用性。
为提高干细胞移植后的存活率并促进恢复，我们展示了成人干细胞——人牙髓干细胞（hDPSCs）和内皮细胞（ECs）在缺氧和条件培养基（CM）作用下三维（3D）自组装形成一种新型球状牙髓类器官。
在培养过程中，通过将原代hDPSCs置于缺氧环境和CM中，诱导其分化为ECs。然后将经缺氧预处理的hDPSCs与ECs按特定比例混合，并在3D环境中进行培养，以产生血管化前的牙髓类器官。分析类器官的生物学特性，并研究与血管生成相关的调控途径。
这两种因子的组合产生了血管化前的人牙髓类器官（V类器官），其在形态和功能上更类似于牙髓组织。整合牙髓组织的单细胞RNA测序和V类器官的RNA测序，以分析与血管生成相关的关键调控途径。确定了叉头框蛋白O1和成纤维细胞生长因子2这两种生物标志物参与V类器官的调控。
在这项创新性研究中，我们有效地建立了V类器官模型，并利用它阐明再生过程中血管生成的新机制，促进临床治疗策略的发展。
