Method for delaying aging caused by in vitro culture of human bone marrow mesenchymal stem cells

Abstract

The invention discloses a method for delaying aging caused by in vitro culture of human bone marrow mesenchymal stem cells; by increasing the expression level of FOXP1 in human MSC cells, the proliferation ability of MSC is improved, the differentiation ability of MSC is protected, and the aging of MSC is delayed . Specifically, lentivirus is used as a vector to transfect human-derived MSCs, and antibiotics are used to screen positive MSC clones and then subculture and expand culture. Compared with the prior art, the present invention can effectively improve the in vitro expansion ability of young-derived MSCs; can reverse the aging symptoms of human-derived MSCs over 70-80 years old; can promote the osteogenic differentiation potential of MSCs; and can inhibit the growth of MSCs. Adipogenic differentiation potential.

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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