Method for delaying senescence due to in vitro culture of human bone marrow MSC (Mesenchymal Stem Cells)

Abstract

The invention discloses a method for delaying senescence due to in vitro culture of human bone marrow MSC (Mesenchymal Stem Cells). By improving the expression level of FOXP1 in the human MSC, the multiplication capability of the MSC is improved, the differentiation capability of the MSC is protected, and the senescence of the MSC is delayed. Concretely, lentiviruses are used as carriers for transfecting the human MSC; and the antibiotic screening positive MSC cloning and passage enlarged culture are performed. Compared with the prior art, the method provided by the invention has the advantages that the in vitro amplification capability of the young human MSC can be effectively improved; the senescence symptom of the old human MSC being older than 70 to 80 years old can be reversed; the osteogenic differentiation potentials of the MSC can be promoted; and the adipogenic differentiation potential of the MSC can be inhibited.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for delaying aging caused by in vitro culture of human bone marrow mesenchymal stem cells (MSC). Background technique [0002] Mesenchymal stem cells (MSCs) are an important type of adult stem cells (Caplan, 1991). MSCs can be detected in multiple parts of body organs, such as umbilical cord blood (Goodwin et al., 2001), among which bone marrow is an important storage site for MSCs (Fehrer and Lepperdinger, 2005; Mendez-Ferret et al., 2010). Bone marrow mesenchymal stem cells have a high ability of self-renewal and the ability to differentiate into osteoblasts, chondrocytes and adipocytes (Bianco et al., 2008a; Bianco et al., 2008b; Nombela-Arrieta et al., 2011; Pittenger et al., 1999), involved in the formation and homeostasis of the skeletal system. Most studies regard osteogenic, adipogenic and chondrogenic differentiation ability as the "gold standard" for d...

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Problems solved by technology

[0017] 1. The use of platelet lysates instead of calf serum will affect the immunosuppressive ability of MSCs and lead to spontaneous differentiation during passage and proliferation of MSCs (Oikonomopoulos et al., 2015)

[0018] 2. Hypoxic culture, 1% O 2 The culture condition of ordinary laboratory is 21% O 2 Concentration, not easy to control and achieve target conditions

[0019] 3. Calorie restriction, Fgf4 protein addition, serum-free medium and other methods are helpful to the in vitro expansion of MSCs, but have no obvious benefit in delaying the aging and differentiation potential of MSCs

[0020] 4. Overexpression of telomerase hTERT in MSCs can easily lead to canceration and spontaneous osteogenic differentiation of MSCs during long-term culture (Gronthos et al., 2003; Simonsen et al., 2002)

[0021] 5. Knocking down the Rb2 gene in MSCs will affect the differentiation potential of MSCs, and their osteogenic and chondrogenic differentiation capabilities will decrease (Alessio et al., 2013)

[0022] 6. Adding Fgf2, PDGF-BB, EGF, vitamin C and other components to the medium is expensive, and long-term subculture will cause MSCs to lose their osteogenic or adipogenic differentiation potential (Coutu et al., 2011; Gharibi and Hughes , 2012)

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