Claims
- 1. An aqueous solution for isolating keratinocytes from human skin, said solution consisting essentially of:
- a) glucose at a concentration of about 10 mM;
- b) N-(2-OH-ethyl-) piperazine -N'-(2-ethanesulfonic acid) at a concentration of 16-22 mM;
- c) sodium chloride at a concentration of 90-140 mM;
- d) potassium chloride at a concentration of about 3 mM;
- e) sodium orthophosphate (Na.sub.2 HPO.sub.4.7H.sub.2 O) at a concentration of 1 mM;
- f) phenol red at a concentration of 0.0033 mM;
- g) about 100 units of penicillin per ml of solution;
- h) about 100 units of streptomycin per ml of solution; and
- i) one component selected from the group consisting of:
- (i) trypsin at a concentration of 0.1%-0.2% w/v; and
- (ii) soy bean trypsin inhibitor at a concentration of 0.1-1.0% w/v.
- 2. The solution of claim 1 wherein said component is soy bean trypsin inhibitor.
- 3. A method for the isolation of basal epidermal keratinocytes from human skin, said method comprising the steps of:
- a) obtaining human skin;
- b) comminuting said skin;
- c) placing said comminuted skin in a solution consisting essentially of:
- i) glucose at a concentration of about 10 mM
- ii) N-(2-OH-ethyl-)piperazine-N'-(2-ethanesulfonic acid) at a concentration of about 16-22 mM
- iii) sodium chloride at a concentration of about 90-140 mM;
- iv) potassium chloride at a concentration of about 3 mM;
- v) sodium orthophosphate at a concentration of about 1 mM;
- vi) phenol red at a concentration of about 0.0033 mM;
- vii) about 100 units of penicillin per mL of solution;
- viii) about 100 units of streptomycin per mL of solution; and
- ix) trypsin at a concentration of 0.1%-0.2% w/v
- at a temperature and for a time sufficient to allow separation of the dermis from the epidermis;
- d) collecting said epidermis; and
- e) collecting said basal keratinocytes from said epidermis in a solution consisting essentially of:
- i) glucose at a concentration of about 10 mM
- ii) N-(2-OH-ethyl-)piperazine-N'-(2-ethanesulfonic acid) at a concentration of about 16-22 mM
- iii) sodium chloride at a concentration of about 90-140 mM;
- iv) potassium chloride at a concentration of about 3 mM;
- v) sodium orthophosphate at a concentration of about 1 mM;
- vi) phenol red at a concentration of about 0.0033 mM;
- vii) about 100 units of penicillin per mL of solution;
- viii) about 100 units of streptomycin per mL of solution; and
- ix) soy bean trypsin at a concentration of 0.1%-1.0% w/v.
- 4. The method of claim 3 wherein said
- a) sodium chloride is a concentration of about 100-130 mM;
- b) N-(2-OH-ethyl-)piperazine-N'-(2-ethanesulfonic acid) is at a concentration of about 18-21 mM;
- c) trypsin is at a concentration of 0.1%-0.18% w/v; and
- d) soy bean trypsin inhibitor at a concentration of 0.3%-0.8% w/v.
- 5. The method of claim 3 wherein said step (c) is conducted for 14-16 hours at a temperature of 4.degree. C.
- 6. The solution of claim 1 wherein said component is trypsin.
- 7. The solution of claim 1 wherein said:
- a) sodium chloride is at a concentration of 100-130 mM;
- b) N-(2-OH-ethyl-) piperazine-N'-(2-ethanesulfonic acid) is at a concentration of 18-21 mM;
- c) trypsin is at a concentration of 0.12-0.18% w/v; and
- d) soy bean trypsin inhibitor is at a concentration of 0.3-0.8% w/v.
Parent Case Info
This application is a continuation of Ser. No. 08/348,569 filed Dec. 2, 1994, now abandoned, which is a continuation of Ser. No. 08/063,872 filed May 18, 1993, now abandoned, which is a continuation of Ser. No. 07/471,976 filed Jan. 29, 1990, now U.S. Pat. No. 5,292,655. The teachings of U.S. Pat. No. 5,292,655 are incorporated herein by reference.
US Referenced Citations (12)
Non-Patent Literature Citations (1)
| Entry |
| Gibco Laboratories, GIBCO Laboratories Catalogue & Reference Guide, pp. 87 and 99, 1987. |
Continuations (3)
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Number |
Date |
Country |
| Parent |
348569 |
Dec 1994 |
|
| Parent |
63872 |
May 1993 |
|
| Parent |
471976 |
Jan 1990 |
|
Patent Grant
5795781
