Compositions and methods for mesenchymal/stromal stem cell rejuvenation and tissue repair by enhanced co-expression of telomerase and myocardin

A composition for generating mammalian stem cells from aging, damaged and/or dysfunctional mammalian cells, wherein said composition comprises:

• rejuvenating senescent mesenchymal stromal cells (MSCs) from a mammalian tissue stroma wherein the rejuvenating MSCs comprise:
• positive expression of telomerase, or a catalytic unit of Telomerase Reverse Transcriptase (TERT);
• and Myocardin (MYCOD);
• positive expression of CD29+, CD44+, and CD90+, and receptors for lipoproteins in said rejuvenating MSCs;
• negative or diminishing expression of CD45, and receptors for acetylated low density lipoprotein (LDL), wherein said rejuvenating senescent MSC's increase at least one of:
• cell regeneration, cell repair, and cell rejuvenation of said damaged cells compared to native MSC's.

A method of making a composition for use in the rejuvenation of aging, damaged and dysfunctional mesenchymal cells, said method comprises:

• isolating a mesenchymal stromal cell (MSC) from an aging, or dysfunctional tissue stroma, wherein said isolated MSCs express low or no TERT and/or MYCOD, and cannot proliferate and differentiate into different mammalian cell lineages in an apolipoprotein-containing medium;
• synthesizing a nuclear acid or cDNA by reverse transcription of mRNA with oligonucleotide primers specific for TERT and MYCOD genes, wherein the cDNA comprises sequences encoding full or a partial length of coding sequences of both TERT and MYCOD genes;
• constructing a expression vector carrying TERT and/or MYCOD cDNA inserts, wherein said TERT/MYCOD-carrying vector comprises a TERT gene and a MYCOD gene;
• and
• transducing said isolated MSCs with a TERT/MYCOD gene-carrying vector;
• and
• forming a rejuvenating MSC, wherein said rejuvenating MSC comprises increased expression of TERT and MYCOD, and thereby comprises therapeutic efficacy for regenerating or repairing diseased mammalian tissues or organs.

A method of making a composition of rejuvenating CD29+/CD44+/CD90+ MSCs, wherein said MSCs are cultivated and harvested in an apolipoprotein-rich medium, said method comprising:

• treating CD29+/CD44+/CD90+ MSCs from adult mammalian tissues with TERT/MYCOD cDNA in a medium comprising ApoAI, ApoB, ApoE and ApoJ, to form TERT/MYCOD-expressing CD29+/CD44+/CD90+ MSCs;
• isolating, and harvesting said isolated TERT/MYCOD-expressing CD29+/CD44+/CD90+ MSCs to form harvested MSCs, wherein said harvested MSCs comprise undifferentiating, non-senescent MSCs;
• and
• separating non-senescent TERT+/MYCOD CD29+/CD44+/CD90+ MSCs from senescent CD29+/CD44+/CD90+ MSCs by passage selection, thereby forming a composition of rejuvenating CD29+/CD44+/CD90+ MSCs.