Application Of Human Induced Pluripotent Stem Cells (hiPSC) In Tissue Engineering And Disease Models

Induced pluripotent stem cells(iPSC)are similar to embryonic stem cells and are a kind of totipotent stem cells with the potential for self-renewal and multi-directional differentiation.hiPSC has a wide range of applications in tissue engineering and disease models.The most significant application is to transform patient adult cells into hiPSC and to induce patient-specific hiPSC to differentiate into mesenchymal stem cells(MSC)to establish disease models.Mechanism research and clinical treatment.However,the traditional hiPSC identification methods such as immunofluorescence based on surface labeling of stem cells and teratoma generation are time-consuming and labor-intensive.The heterogeneity between different hiPSC clones affects the differentiation results,and the small molecule induces a long differentiation cycle and many factors.Therefore,how to efficiently identify high-purity hiPSC in vitro and differentiate them into functional hiPSC-MSC in large quantities remains challenging.This study combines artificial intelligence and deep machine learning to establish a new hiPSC clone quality evaluation method,and at the same time establish a simple and efficient method to induce differentiated hiPSC to obtain a large number of functional MSC.Based on the method established above,an hiPSC disease model was established for Chinese patients with premature senility(HGPS)and induced to differentiate into patient-specific hiPSC-MSC,and the disease mechanism was initially explored.First,we used the hiPSC reprogramming system of Sendai virus to reprogram healthy human fibroblasts to obtain hiPSC samples of various quality.Then,the bright field images of the microscope cloned by hiPSC were collected and the data was marked,and a multi-source feature ensemble learning(MsFEL)model was established.Finally,the data is biologically verified.The established evaluation model was used to select multiple hiPSC of good quality to be cultured in a stem cell system without mouse trophoblasts.The optimized MSC differentiation medium was used to replace the stem cell culture medium to induce hiPSC to differentiate into MSC.Subsequently,hiPSC-MSC was subjected to surface antigen analysis and in vitro multi-directional differentiation ability identification,and the obtained hiPSC-MSC was used to repair SD rat cartilage injury in vivo.Finally,we established hiPSC cell lines of 2 Chinese premature aging in the same family by reprogramming and used the established evaluation model to select hiPSC clones and conduct MSC differentiation at the same time.In this paper,15700 hiPSC clones were identified by artificial intelligence,which achieved a classification accuracy of 95.55%,established hiPSC clone quality evaluation standards,and was verified by live cell staining.We use a simple one-step method to successfully induce and differentiate hiPSC into functional hiPSC-MSC.hiPSC-MSC showed a similar surface antigen pattern(CD73,CD90,CD105 positive and CD34,CD45 negative)as well as osteogenic,chondrogenic and adipogenic differentiation ability to those derived from bone marrow.The SD rat cartilage injury repair experiment shows that the hiPSC-MSC obtained by our differentiation has the ability to repair in vivo.In addition,using the hiPSC clone identification and hiPSC-MSC generation method established in this thesis,we successfully established hiPSC cell lines of 2 Chinese premature aging in the same family through reprogramming.The mutation site verification proved that the patient's hiPSC carried a homozygous R527 C mutation,and the pluripotency test showed that the mutation did not change the hiPSC's pluripotency expression.After hiPSC-MSC differentiation,CD90 abnormality appeared,suggesting that its pathogenic mechanism may be related to the regulation of CD90 expression.