Immortal human bone marrow matrix stem cell line and its establishing process

Abstract

The present invention is immortal human bone marrow matrix stem cell line and its establishing process. The primary human bone marrow matrix stem cell from normally aborted fetus bone marrow tissue is single cell cloned and cultured in a finite dilution process to purify primary cell and transfected with pCIneo-hTERT gene for immortalizing. The human bone marrow matrix stem cell line is extracorporeal cultured, may be infinitely cloned and proliferated and is normal cytologic cell without conversion cell or cancer cell characteristic. The cell may be directionally induced and differentiated into bone cell, cartilage cell or cardiac muscle-like cell and has multiple direction differentiating latent and adult stem cell characteristic. It may be applied in biological engineering field. The present invention provides standard cell line for the biological research of biological behavior of human bone marrow matrix stem cell, platform for basic tissue engineering research, enough seed cells for clinical application and effective method for relevant research.

Description

technical field [0001] The invention belongs to the field of human and mammalian cell (tissue) engineering in the subject of biotechnology, and relates to in vitro separation, purification, identification, gene transfection, immortalization and multidirectional induction of human bone marrow stromal stem cells, in particular to an immortalized human bone marrow stromal stem cell system and how to create it. Background technique [0002] Due to certain factors such as trauma, infection, tumor, etc., the bone loses some or a section of bone, forming a large gap, which is called a bone defect. Most of the bone defects are difficult to heal, and finally form a nonunion. Due to the large defect gap, it is difficult for osteoblasts to crawl through the gap and the normal healing process cannot occur, and it is only filled by fibrous tissue. For this reason, many scholars have carried out relevant research and obtained some achievements, but for large...

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Problems solved by technology

[0006] (3) The interaction between BMSCs, the interaction with hematopoietic stem cells, and the interaction with cells at different mature stages directly affect their growth and differentiation, but the mechanism of these effects is still unclear

[0008] (5) There is a problem of cell aging in BMSCs culture in vitro, which limits the number of available cells and cannot meet the requirements of tissue engineering

[0010] To sum up, human bone marrow stromal stem cells have not been fully characterized at present, and its isolation, purification and expansion methods are still being explored, and the identification results of human bone marrow stromal stem cells obtained are not consistent. Report on Human Bone Marrow Stromal Stem Cell Line

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